Compounds for the Treatment of Alzheimer&#39;s Disease

ABSTRACT

The present disclosure provides compounds that are amylin receptor antagonist compounds, compositions that include the subject compounds, methods for preparing and using the amylin receptor antagonists, and compositions containing the amylin receptor antagonists for treating, preventing, or ameliorating Alzheimer&#39;s disease. Aspects of the present disclosure include a method of inhibiting activity of an amylin receptor by administering to a subject in need thereof a therapeutically effective amount of an amylin receptor antagonist.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.63/094,777, filed Oct. 21, 2020, the disclosure of which is incorporatedherein by reference.

INTRODUCTION

Alzheimer's disease is the most common form of dementia that ischaracterized by deposition of amyloid β-protein (Aβ or Abeta) intra-and extracellularly within cortical and limbic brain structures criticalfor memory and cognitive functions (Selkoe, 1994 and 2013; Hardy et al.,2002). A central question in Alzheimer's disease research is whether theamyloid protein is a cause or a consequence of the disease. Presently,it appears that the likely answer is both (Hardy, 2009). Evidencestrongly supports a role for Aβ in the pathogenesis of Alzheimer'sdisease, namely: a) Alzheimer's disease associated with inheritedAmyloid Precursor Protein (APP) mutations; b) neurotoxicity of solubleoligomeric Aβ when applied to neurons; and c) APP overexpressing micethat recapitulate certain neuropathological and behavioral features ofAlzheimer's disease (Liu et al., 2012; Bateman et al., 2012; Patel etal., 2012; Danysz et al., 2012). On the other hand, adverse events inclinical trials for Alzheimer's disease using Aβ vaccine-based therapy,and the subsequent failure of monoclonal antibody therapies andinhibitors of the Aβ generating gamma-secretase enzyme in improvingcognitive functions in patients have forced reconsideration of theseapproaches as disease-modifying treatment strategies in Alzheimer'sdisease (Liu et al., 2012). Nonetheless, it is hard to imagine adefinitive treatment that will not serve to ameliorate in some form theneurotoxic effects of Aβ, since this is a key “upstream” event inAlzheimer's disease pathogenesis (as established by alterations in CSFAβ levels decades before clinical onset) (Bateman et al., 2012).

Multiple receptors have been implicated in mediating Aβ disruption ofneuronal and synaptic processes in Alzheimer's disease, and thusidentified as potential targets for developing anti-Aβ therapies (Patelet al., 2012; Danysz et al., 2012). The amylin receptor, comprised ofheterodimers of the calcitonin receptor with receptor activity-modifyingproteins, serves as a portal for the expression of deleterious effectsof Aβ and human amylin (Fu et al., 2012). Amylin is a 37-amino acidpeptide hormone that is co-secreted with insulin by beta cells of thepancreas that control glucose levels in blood.

Both Aβ and human amylin are amyloidogenic peptides which sharestructure-functional relationships; for example, both peptides aggregateand form soluble and insoluble oligomeric intermediates. Amylin has thepropensity to aggregate and form amyloid oligomers and fibrils in thepancreas in type 2 diabetes (Westermark et al., 2011) and in Alzheimer'sdisease brains (Abedini et al., 2013). Aβ and human amylin causedysfunction and death of neurons preferentially affected in Alzheimer'sdisease (Jhamandas et al., 2011; 2004). Neurotoxic effects of humanamylin and Aβ are expressed through the amylin receptor 3 subtype(AMY3).

Amylin receptor antagonists, such as AC253 (a 24-amino acid peptide),are neuroprotective against Aβ toxicity (Jhamandas et al., 2004; 2011;2012). Down-regulation of amylin receptor gene expression using siRNAmitigates oligomerized Aβ-induced toxicity (Jhamandas et al., 2011). InAlzheimer's disease transgenic model mice (TgCRND8) which over-expressAβ, amylin receptor was up-regulated within specific brain regions thatdemonstrate an increased burden of amyloid beta deposits (Jhamandas etal., 2011). Blockade of the amylin receptor with AC253 can reverseimpairment of Aβ- or human amylin-induced depression of long-termpotentiation, a cellular surrogate of memory, as observed in thehippocampus of Alzheimer's disease mice (TgCRND8) (Kimura et al., 2012).Similar benefits have been reported with pramlintide, a syntheticnon-amyloidogenic analog of amylin. While data support a neuroprotectiverole for this compound, it appears to act as an amylin receptorantagonist rather than an agonist (Kimura et al., 2016). Although amylinreceptor antagonist AC253 peptide has therapeutic potential inAlzheimer's disease, it suffers from poor enzymatic stability and aninability to penetrate the blood brain barrier.

SUMMARY

The present disclosure provides compounds that are amylin receptorantagonists, compositions that include the subject compounds, andmethods for preparing and using the amylin receptor antagonists and thecompositions for treating, preventing, or ameliorating Alzheimer'sdisease.

Aspects of the present disclosure include a method of inhibitingactivity of an amylin receptor. The method includes administering to asubject in need thereof, a therapeutically effective amount of acompound of formula (I):

wherein:

R is selected from the group consisting of —H, C₁-C₆-alkyl, andsubstituted C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H,C₁-C₆-alkyl, and substituted C₁-C₆-alkyl;

each W is selected from the group consisting of —CH₂—, —CHR³— and—CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, substituted carbocycle or oxo;

m is selected from 1, 2 or 3;

Q is selected from the group consisting of —CH₂—, —C(═O)—, —CH₂C(═O)—,and —CH₂CH₂—;

X is present or absent, and if present is selected from the groupconsisting of —CH₂— and —C(═O)—;

Y is selected from the group consisting of —CH₂—, and —C(═O)—;

Z is selected from the group consisting of —OR⁵, halogen, —CN, —NR⁵R⁵,—NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and

each R⁵ is independently selected from the group consisting of —H,C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, the amylin receptor is an AMY3 receptor.

In certain embodiments, the administering is effective for treating adisease mediated through activity of the amylin receptor. In certainembodiments, the disease is Alzheimer's disease.

In certain embodiments, the compound is of formula (II):

wherein:

R is selected from the group consisting of —H and C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H, andC₁-C₆-alkyl;

each W is selected from the group consisting of —CH₂—, —CHR³— and—CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, substituted carbocycle or oxo;

m is selected from 1, 2 or 3;

Q is selected from the group consisting of —CH₂—, —C(═O)— and—CH₂C(═O)—;

Y is selected from the group consisting of —CH₂—, and —C(═O)—;

Z is selected from the group consisting of —OR⁵, -halogen, —NR⁵R⁵,—NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and

each R⁵ is independently selected from the group consisting of —H,C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, the compound is of formula (III):

wherein:

R is selected from the group consisting of —H and C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H, andC₁-C₆-alkyl;

each W is selected from the group consisting of —CH₂—, —CHR³— and—CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, substituted carbocycle or oxo;

m is selected from 1, 2 or 3;

Z is selected from the group consisting of —OR⁵, halogen, —NR⁵R⁵,—NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and

each R⁵ is independently selected from the group consisting of —H,C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, R is —H or —CH₃.

In certain embodiments, each R¹ is selected from —H, —CH₃, —F, —Cl,—CF₃, and heteroaryl.

In certain embodiments, each R¹ is selected from —H, —CH₃, —F, —Cl,—CF₃, and pyridyl.

In certain embodiments, R¹ is —H.

In certain embodiments, m is 1. In certain embodiments, m is 2.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heteroaryl, substitutedheteroaryl, fused-heterocycle, and substituted fused-heterocycle, ortogether R³ and R⁴ comprise a C₃-C₆ carbocycle, substituted C₃-C₆carbocycle or oxo.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of —F, —Cl, —CH₃, or together R³ and R⁴ comprise acyclopropyl, cyclobutyl, cyclopentyl or oxo.

In certain embodiments, W is selected from —CH₂— and —CHR³—.

In certain embodiments, W is —CH₂—.

In certain embodiments, Z is selected from the group consisting of —OR⁵,—F, —NHC(═O)R⁵, and heteroaryl.

In certain embodiments, Z is selected from the group consisting of —OR⁵,and —NHC(═O)R⁵.

In certain embodiments, each R⁵ is independently selected from —H, —CH₃,—CH₂CH₃, —CH(CH₃)₂ and phenyl.

In certain embodiments, each R⁵ is independently selected from —H, and—CH₃.

In certain embodiments, the compound is selected from the groupconsisting of:

Aspects of the present disclosure include a compound of formula (IV):

wherein:

R is selected from the group consisting of —H and C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H, andC₁-C₆-alkyl;

each W is independently selected from the group consisting of —CH₂—,—CHR³— and —CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, or substituted carbocycle;

Z is selected from the group consisting of —OH, —OC(═O)CH₃, —OC(═O)Ph,and —NHC(═O)CH₃;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof; or a pharmaceutically acceptable salt,solvate, hydrate, or prodrug thereof;

with the proviso that the compound is not:

N-[3-(3,4-dihydro-3-oxo-1(2H)-quinoxalinyl)-3-oxopropyl]acetamide.

In certain embodiments, R is —CH₂CH₃ or —CH₃.

In certain embodiments, R is —H.

In certain embodiments, each R¹ is independently selected from the groupconsisting of —H, —F, —Cl, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, —OCH₃,heteroaryl, and substituted heteroaryl.

In certain embodiments, each R¹ is independently selected from the groupconsisting of —H, —F, —Cl, —CH₃, —CF₃, —OCH₃, and pyridyl.

In certain embodiments, each R¹ is —H.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of —F, —Cl, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, or together R³ and R⁴ comprise aC₃-C₆-cycloalkyl, or substituted C₃-C₆-cycloalkyl.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of —F, —CH₃, —CH₂CH₃, or together R³ and R⁴ comprisecyclopropyl, substituted cyclopropyl, cyclobutyl, substitutedcyclobutyl, cyclopentyl, or substituted cyclopentyl.

In certain embodiments, each W is selected from —CHR³— and —CR³R⁴—.

In certain embodiments, each W is —CH₂—.

In certain embodiments, Z is selected from the group consisting of —OH,and —OC(═O)CH₃.

In certain embodiments, Z is —OH.

In certain embodiments, the compound is selected from:

Aspects of the present disclosure include a compound of Formula (V),Formula (VI), or Formula (VII):

wherein:

R⁷ is selected from the group consisting of —CH₂OC(═O)OR⁸,—C(CH₃)HOC(═O)OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NHR⁸, —C(═O)NR⁸R⁸, andC₁-C₆-alkyl; and

each R⁸ is independently selected from C₁-C₆-alkyl, substitutedC₂-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,heterocycyl, substituted heterocycyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl,

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, the compound is selected from:

Aspects of the present disclosure include a method of inhibitingactivity of an amylin receptor, where the method includes administeringto a subject in need thereof, a therapeutically effective amount of acompound of formula (IV), formula (V), formula (VI), or formula (VII) ofthe present disclosure.

In certain embodiments, the amylin receptor is an AMY3 receptor.

In certain embodiments, the administering is effective for treating adisease mediated through activity of the amylin receptor. In certainembodiments, the disease is Alzheimer's disease.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Screening of compounds using ELISA assay for cAMP in AMY3transfected cells. cAMP generation by 0.5 μM was normalized to 100% andthe ability of individual compounds to attenuate this response in thepresence of human amylin was assessed.

FIG. 2: Human amylin (0.1 μM) generation of cAMP was attenuated in thepresence of Compounds 1 and 10 (10 μM)

FIG. 3: Compound 1 and 10 (0.01-10 μM range) attenuated elevations inintracellular calcium evoked by application of human amylin (0.5 μM).Fluor-8 is a fluorescent membrane permeable calcium dye that fluorescesupon elevation of intracellular calcium levels due to ligand activationof receptors (such as human amylin).

FIG. 4: Compounds 1 and 10 were cytoprotective against amyloid beta(Abeta) toxicity in a neurons cell line (N2a). MTT cell assay measuredcell viability. Abeta was prepared and applied as a soluble oligomericspecies described in the literature, such as in Soudy, R. et alAlzheimer's & Dementia: Translational Research & Clinical Interventions(2017).

FIGS. 5A to 5C: LTP data for Compound 1, which blocked human Amylin andAmyloid beta induced LTP responses. Compound 1 also restored LTP intransgenic (Tg_Cont) AD mice to levels comparable to those inage-matched wild type controls.

FIGS. 6A to 6C: Histograms depicting summary data of LTP responses fromexperimental data shown in FIG. 5. Right panels show statisticaltreatment of the data using one-way ANOVA followed Tukey's test. Bothhuman amylin- and Aβ-induced LTP responses were blocked by Compound 1and the chronically reduced LTP levels in transgenic transgenic AD mice(Tg_Cont) were restored by applications of Compound 1 to levelscomparable to age matched wild type control mice (WT_Cont). Compound 1had no effects on LTP from WT_Cont mice.

FIGS. 7A to 7C: LTP data for Compound 10, which blocked human Amylin andAmyloid beta induced LTP responses. Compound 10 also restored LTP intransgenic (Tg_Cont) AD mice to levels comparable to those inage-matched wild type controls.

FIGS. 8A to 8C: Histograms depicting summary data of LTP responses fromexperimental data shown in FIG. 5. Right panels show statisticaltreatment of the data using one-way ANOVA followed Tukey's test. Bothhuman amylin- and Aβ-induced LTP responses were blocked by Compound 10and the chronically reduced LTP levels in transgenic transgenic AD mice(Tg_Cont) were restored by applications of Compound 10 to levelscomparable to age matched wild type control mice (WT_Cont). Compound 10had no effects on LTP from WT_Cont mice.

DEFINITIONS

The following terms have the following meanings unless otherwiseindicated. Any undefined terms have their art recognized meanings.

“Alkyl” refers to monovalent saturated aliphatic hydrocarbyl groupshaving from 1 to 10 carbon atoms and such as 1 to 6 carbon atoms, or 1to 5, or 1 to 4, or 1 to 3 carbon atoms. This term includes, by way ofexample, linear and branched hydrocarbyl groups such as methyl (CH₃—),ethyl (CH₃CH₂—), n-propyl (CH₃CH₂CH₂—), isopropyl ((CH₃)₂CH—), n-butyl(CH₃CH₂CH₂CH₂—), isobutyl ((CH₃)₂CHCH₂—), sec-butyl ((CH₃)(CH₃CH₂)CH—),t-butyl ((CH₃)₃C—), n-pentyl (CH₃CH₂CH₂CH₂CH₂—), and neopentyl((CH₃)₃CCH₂—).

The term “substituted alkyl” refers to an alkyl group as defined hereinwherein one or more carbon atoms in the alkyl chain (except the C₁carbon atom) have been optionally replaced with a heteroatom such as—O—, —N—, —S—, —S(O)_(n)— (where n is 0 to 2), —NR— (where R is hydrogenor alkyl) and having from 1 to 5 substituents selected from the groupconsisting of alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano,halogen, hydroxyl, oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy,thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substitutedthioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-aryl,—SO-heteroaryl, —SO₂-alkyl, —SO₂-aryl, —SO₂-heteroaryl, and—NR^(a)R^(b), wherein R^(a) and R^(b) may be the same or different andare chosen from hydrogen, optionally substituted alkyl, cycloalkyl,alkenyl, cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic.

“Alkylene” refers to divalent aliphatic hydrocarbyl groups preferablyhaving from 1 to 6 and more preferably 1 to 3 carbon atoms that areeither straight-chained or branched, and which are optionallyinterrupted with one or more groups selected from —O—, —NR¹⁰—,—NR¹⁰C(O)—, —C(O)NR¹⁰— and the like, where R¹⁰ is chosen from chosenfrom hydrogen, optionally substituted alkyl, cycloalkyl, alkenyl,cycloalkenyl, alkynyl, aryl, heteroaryl and heterocyclic. This termincludes, by way of example, methylene (—CH₂—), ethylene (—CH₂CH₂—),n-propylene (—CH₂CH₂CH₂—), iso-propylene (—CH₂CH(CH₃)—),(—C(CH₃)₂CH₂CH₂—), (—C(CH₃)₂CH₂C(O)—), (—C(CH₃)₂CH₂C(O)NH—),(—CH(CH₃)CH₂—), and the like.

“Substituted alkylene” refers to an alkylene group having from 1 to 3hydrogens replaced with substituents as described for carbons in thedefinition of “substituted” below.

The term “alkane” refers to alkyl group and alkylene group, as definedherein.

The term “alkylaminoalkyl”, “alkylaminoalkenyl” and “alkylaminoalkynyl”refers to the groups R′NHR″— where R′ is alkyl group as defined hereinand R″ is alkylene, alkenylene or alkynylene group as defined herein.

The term “alkaryl” or “aralkyl” refers to the groups -alkylene-aryl and-substituted alkylene-aryl where alkylene, substituted alkylene and arylare defined herein.

“Alkoxy” refers to the group —O-alkyl, wherein alkyl is as definedherein. Alkoxy includes, by way of example, methoxy, ethoxy, n-propoxy,isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, and the like. Theterm “alkoxy” also refers to the groups alkenyl-O—, cycloalkyl-O—,cycloalkenyl-O—, and alkynyl-O—, where alkenyl, cycloalkyl,cycloalkenyl, and alkynyl are as defined herein.

The term “substituted alkoxy” refers to the groups substituted alkyl-O—,substituted alkenyl-O—, substituted cycloalkyl-O—, substitutedcycloalkenyl-O—, and substituted alkynyl-O— where substituted alkyl,substituted alkenyl, substituted cycloalkyl, substituted cycloalkenyland substituted alkynyl are as defined herein.

The term “alkoxyamino” refers to the group —NH-alkoxy, wherein alkoxy isdefined herein.

The term “haloalkoxy” refers to the groups alkyl-O— wherein one or morehydrogen atoms on the alkyl group have been substituted with a halogroup and include, by way of examples, groups such as trifluoromethoxy,and the like.

The term “haloalkyl” refers to a substituted alkyl group as describedabove, wherein one or more hydrogen atoms on the alkyl group have beensubstituted with a halo group. Examples of such groups include, withoutlimitation, fluoroalkyl groups, such as trifluoromethyl, difluoromethyl,trifluoroethyl and the like.

The term “alkylalkoxy” refers to the groups -alkylene-O-alkyl,alkylene-O-substituted alkyl, substituted alkylene-O-alkyl, andsubstituted alkylene-O-substituted alkyl wherein alkyl, substitutedalkyl, alkylene and substituted alkylene are as defined herein.

“Alkenyl” refers to straight chain or branched hydrocarbyl groups havingfrom 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and havingat least 1 and preferably from 1 to 2 sites of double bond unsaturation.This term includes, by way of example, bi-vinyl, allyl, andbut-3-en-1-yl. Included within this term are the cis and trans isomersor mixtures of these isomers.

The term “substituted alkenyl” refers to an alkenyl group as definedherein having from 1 to 5 substituents, or from 1 to 3 substituents,selected from alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO— substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

“Alkynyl” refers to straight or branched monovalent hydrocarbyl groupshaving from 2 to 6 carbon atoms and preferably 2 to 3 carbon atoms andhaving at least 1 and preferably from 1 to 2 sites of triple bondunsaturation. Examples of such alkynyl groups include acetylenyl(—C≡CH), and propargyl (—CH₂C≡CH).

The term “substituted alkynyl” refers to an alkynyl group as definedherein having from 1 to 5 substituents, or from 1 to 3 substituents,selected from alkoxy, substituted alkoxy, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl, acylamino,acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO— substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl, and —SO₂-heteroaryl.

“Alkynyloxy” refers to the group —O-alkynyl, wherein alkynyl is asdefined herein. Alkynyloxy includes, by way of example, ethynyloxy,propynyloxy, and the like.

“Acyl” refers to the groups H—C(O)—, alkyl-C(O)—, substitutedalkyl-C(O)—, alkenyl-C(O)—, substituted alkenyl-C(O)—, alkynyl-C(O)—,substituted alkynyl-C(O)—, cycloalkyl-C(O)—, substitutedcycloalkyl-C(O)—, cycloalkenyl-C(O)—, substituted cycloalkenyl-C(O)—,aryl-C(O)—, substituted aryl-C(O)—, heteroaryl-C(O)—, substitutedheteroaryl-C(O)—, heterocyclyl-C(O)—, and substitutedheterocyclyl-C(O)—, wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein. For example, acylincludes the “acetyl” group CH₃C(O)—

“Acylamino” refers to the groups —NR²⁰C(O)alkyl, —NR²⁰C(O)substitutedalkyl, NR²⁰C(O)cycloalkyl, —NR²⁰C(O)substituted cycloalkyl,—NR²⁰C(O)cycloalkenyl, —NR²⁰C(O)substituted cycloalkenyl,—NR²⁰C(O)alkenyl, —NR²⁰C(O)substituted alkenyl, —NR²⁰C(O)alkynyl,—NR²⁰C(O)substituted alkynyl, —NR²⁰C(O)aryl, —NR²⁰C(O)substituted aryl,—NR²⁰C(O)heteroaryl, —NR²⁰C(O)substituted heteroaryl,—NR²⁰C(O)heterocyclic, and —NR²⁰C(O)substituted heterocyclic, whereinR²⁰ is hydrogen or alkyl and wherein alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Aminocarbonyl” or the term “aminoacyl” refers to the group—C(O)NR²¹R²², wherein R²¹ and R²² independently are selected from thegroup consisting of hydrogen, alkyl, substituted alkyl, alkenyl,substituted alkenyl, alkynyl, substituted alkynyl, aryl, substitutedaryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic and where R²¹ and R²² are optionally joinedtogether with the nitrogen bound thereto to form a heterocyclic orsubstituted heterocyclic group, and wherein alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic, andsubstituted heterocyclic are as defined herein.

“Aminocarbonylamino” refers to the group —NR²¹C(O)NR²²R²³ where R²¹,R²², and R²³ are independently selected from hydrogen, alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic and where R²¹ andR²² are optionally joined together with the nitrogen bound thereto toform a heterocyclic or substituted heterocyclic group, and whereinalkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

The term “alkoxycarbonylamino” refers to the group —NR^(d)C(O)OR^(d)where each R^(d) is independently hydrogen, alkyl, substituted alkyl,aryl, heteroaryl, or heterocyclyl wherein alkyl, substituted alkyl,aryl, heteroaryl, and heterocyclyl are as defined herein.

The term “acyloxy” refers to the groups alkyl-C(O)O—, substitutedalkyl-C(O)O—, cycloalkyl-C(O)O—, substituted cycloalkyl-C(O)O—,aryl-C(O)O—, heteroaryl-C(O)O—, and heterocyclyl-C(O)O— wherein alkyl,substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, heteroaryl,and heterocyclyl are as defined herein.

“Aminosulfonyl” refers to the group —SO₂NR²¹R²², wherein R²¹ and R²²independently are selected from hydrogen, alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl,substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, heteroaryl, substituted heteroaryl,heterocyclic, and substituted heterocyclic and where R²¹ and R²² areoptionally joined together with the nitrogen bound thereto to form aheterocyclic or substituted heterocyclic group, and wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic are as definedherein.

“Sulfonylamino” refers to the group —NR²¹SO₂R²², wherein R²¹ and R²²independently are selected from hydrogen, alkyl, substituted alkyl,alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl,substituted aryl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, heteroaryl, substituted heteroaryl,heterocyclic, and substituted heterocyclic and where R²¹ and R²² areoptionally joined together with the nitrogen bound thereto to form aheterocyclic or substituted heterocyclic group, and wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic are as definedherein.

“Aryl” or “Ar” refers to a monovalent aromatic carbocyclic group of from6 to 18 carbon atoms having a single ring (such as is present in aphenyl group) or a ring system having multiple condensed rings (examplesof such aromatic ring systems include naphthyl, anthryl and indanyl)which condensed rings may or may not be aromatic, provided that thepoint of attachment is through an atom of an aromatic ring. This termincludes, by way of example, phenyl and naphthyl. Unless otherwiseconstrained by the definition for the aryl substituent, such aryl groupscan optionally be substituted with from 1 to 5 substituents, or from 1to 3 substituents, selected from acyloxy, hydroxy, thiol, acyl, alkyl,alkoxy, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl,substituted alkoxy, substituted alkenyl, substituted alkynyl,substituted cycloalkyl, substituted cycloalkenyl, amino, substitutedamino, aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl,carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy,heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl, —SO₂-heteroaryl and trihalomethyl.

“Aryloxy” refers to the group —O-aryl, wherein aryl is as definedherein, including, by way of example, phenoxy, naphthoxy, and the like,including optionally substituted aryl groups as also defined herein.

“Amino” refers to the group —NH₂.

The term “substituted amino” refers to the group —NR^(m)R^(m) where eachR^(m) is independently selected from the group consisting of hydrogen,alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl,substituted alkenyl, cycloalkenyl, substituted cycloalkenyl, alkynyl,substituted alkynyl, aryl, heteroaryl, and heterocyclyl provided that atleast one R is not hydrogen.

The term “azido” refers to the group —N₃.

“Carboxyl,” “carboxy” or “carboxylate” refers to —CO₂H or salts thereof.

“Carboxyl ester” or “carboxy ester” or the terms “carboxyalkyl” or“carboxylalkyl” refers to the groups —C(O)O-alkyl, —C(O)O-substitutedalkyl, —C(O)O-alkenyl, —C(O)O-substituted alkenyl, —C(O)O-alkynyl,—C(O)O-substituted alkynyl, —C(O)O-aryl, —C(O)O-substituted aryl,—C(O)O-cycloalkyl, —C(O)O-substituted cycloalkyl, —C(O)O-cycloalkenyl,—C(O)O-substituted cycloalkenyl, —C(O)O-heteroaryl, —C(O)O-substitutedheteroaryl, —C(O)O-heterocyclic, and —C(O)O-substituted heterocyclic,wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl,substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl,substituted cycloalkenyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, heterocyclic, and substituted heterocyclic areas defined herein.

“(Carboxyl ester)oxy” or “carbonate” refers to the groups —O—C(O)O—alkyl, —O—C(O)O-substituted alkyl, —O—C(O)O-alkenyl,—O—C(O)O-substituted alkenyl, —O—C(O)O-alkynyl, —O—C(O)O-substitutedalkynyl, —O—C(O)O-aryl, —O—C(O)O-substituted aryl, —O—C(O)O-cycloalkyl,—O—C(O)O-substituted cycloalkyl, —O—C(O)O-cycloalkenyl, —O—C(O)O—substituted cycloalkenyl, —O—C(O)O-heteroaryl, —O—C(O)O-substitutedheteroaryl, —O—C(O)O-heterocyclic, and —O—C(O)O-substitutedheterocyclic, wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic are as defined herein.

“Cyano” or “nitrile” refers to the group —CN.

“Cycloalkyl” refers to cyclic alkyl groups of from 3 to 10 carbon atomshaving single or multiple cyclic rings including fused, bridged, andspiro ring systems. Examples of suitable cycloalkyl groups include, forinstance, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyland the like. Such cycloalkyl groups include, by way of example, singlering structures such as cyclopropyl, cyclobutyl, cyclopentyl,cyclooctyl, and the like, or multiple ring structures such asadamantanyl, and the like.

The term “substituted cycloalkyl” refers to cycloalkyl groups havingfrom 1 to 5 substituents, or from 1 to 3 substituents, selected fromalkyl, substituted alkyl, alkoxy, substituted alkoxy, cycloalkyl,substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, acyl,acylamino, acyloxy, amino, substituted amino, aminoacyl, aminoacyloxy,oxyaminoacyl, azido, cyano, halogen, hydroxyl, oxo, thioketo, carboxyl,carboxylalkyl, thioaryloxy, thioheteroaryloxy, thioheterocyclooxy,thiol, thioalkoxy, substituted thioalkoxy, aryl, aryloxy, heteroaryl,heteroaryloxy, heterocyclyl, heterocyclooxy, hydroxyamino, alkoxyamino,nitro, —SO-alkyl, —SO-substituted alkyl, —SO-aryl, —SO-heteroaryl,—SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

“Cycloalkenyl” refers to non-aromatic cyclic alkyl groups of from 3 to10 carbon atoms having single or multiple rings and having at least onedouble bond and preferably from 1 to 2 double bonds.

The term “substituted cycloalkenyl” refers to cycloalkenyl groups havingfrom 1 to 5 substituents, or from 1 to 3 substituents, selected fromalkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl,cycloalkenyl, substituted cycloalkenyl, acyl, acylamino, acyloxy, amino,substituted amino, aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano,halogen, hydroxyl, keto, thioketo, carboxyl, carboxylalkyl, thioaryloxy,thioheteroaryloxy, thioheterocyclooxy, thiol, thioalkoxy, substitutedthioalkoxy, aryl, aryloxy, heteroaryl, heteroaryloxy, heterocyclyl,heterocyclooxy, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl.

“Cycloalkynyl” refers to non-aromatic cycloalkyl groups of from 5 to 10carbon atoms having single or multiple rings and having at least onetriple bond.

“Carbocycle” refers to non-aromatic or aromatic cyclic groups, such ascycloalkyl, cycloalkenyl, cycloalkynyl, and aryl groups as definedherein. A carbocycle goup may be unsubstituted or substituted as definedherein.

“Cycloalkoxy” refers to —O-cycloalkyl.

“Cycloalkenyloxy” refers to —O-cycloalkenyl.

“Halo” or “halogen” refers to fluoro, chloro, bromo, and iodo.

“Hydroxy” or “hydroxyl” refers to the group —OH.

“Heteroaryl” refers to an aromatic group of from 1 to 15 carbon atoms,such as from 1 to 10 carbon atoms and 1 to 10 heteroatoms selected fromthe group consisting of oxygen, nitrogen, and sulfur within the ring.Such heteroaryl groups can have a single ring (such as, pyridinyl,imidazolyl or furyl) or multiple condensed rings in a ring system (forexample as in groups such as, indolizinyl, quinolinyl, benzofuran,benzimidazolyl or benzothienyl), wherein at least one ring within thering system is aromatic. To satisfy valence requirements, anyheteroatoms in such heteroaryl rings may or may not be bonded to H or asubstituent group, e.g., an alkyl group or other substituent asdescribed herein. In certain embodiments, the nitrogen and/or sulfurring atom(s) of the heteroaryl group are optionally oxidized to providefor the N-oxide (N→O), sulfinyl, or sulfonyl moieties. This termincludes, by way of example, pyridinyl, pyrrolyl, indolyl, thiophenyl,and furanyl. Unless otherwise constrained by the definition for theheteroaryl substituent, such heteroaryl groups can be optionallysubstituted with 1 to 5 substituents, or from 1 to 3 substituents,selected from acyloxy, hydroxy, thiol, acyl, alkyl, alkoxy, alkenyl,alkynyl, cycloalkyl, cycloalkenyl, substituted alkyl, substitutedalkoxy, substituted alkenyl, substituted alkynyl, substitutedcycloalkyl, substituted cycloalkenyl, amino, substituted amino,aminoacyl, acylamino, alkaryl, aryl, aryloxy, azido, carboxyl,carboxylalkyl, cyano, halogen, nitro, heteroaryl, heteroaryloxy,heterocyclyl, heterocyclooxy, aminoacyloxy, oxyacylamino, thioalkoxy,substituted thioalkoxy, thioaryloxy, thioheteroaryloxy, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl, andtrihalomethyl.

The term “heteroaralkyl” refers to the groups -alkylene-heteroaryl wherealkylene and heteroaryl are defined herein. This term includes, by wayof example, pyridylmethyl, pyridylethyl, indolylmethyl, and the like.

“Heteroaryloxy” refers to —O-heteroaryl.

“Heterocycle,” “heterocyclic,” “heterocycloalkyl,” and “heterocyclyl”refer to a saturated or unsaturated group having a single ring ormultiple condensed rings, including fused bridged and spiro ringsystems, and having from 3 to 20 ring atoms, including 1 to 10 heteroatoms. These ring atoms are selected from nitrogen, sulfur, or oxygen,where, in fused ring systems, one or more of the rings can becycloalkyl, heterocyclyl, aryl, or heteroaryl, provided that the pointof attachment is through the non-aromatic ring. Fused ring systemsinclude compounds where two rings share two adjacent atoms. In fusedheterocycle systems one or both of the two fused rings can beheterocyclyl. In certain embodiments, the nitrogen and/or sulfur atom(s)of the heterocyclic group are optionally oxidized to provide for theN-oxide, —S(O)—, or —SO₂— moieties. To satisfy valence requirements, anyheteroatoms in such heterocyclic rings may or may not be bonded to oneor more H or one or more substituent group(s), e.g., an alkyl group orother substituent as described herein.

Examples of heterocycles and heteroaryls include, but are not limitedto, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole,indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine,naphthylpyridine, quinoxaline, 1,2,3,4-tetrahydroquinoxaline,quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine,acridine, phenanthroline, isothiazole, phenazine, isoxazole,phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine,piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline,4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene,benzo[b]thiophene, morpholinyl, 3,4-dihydro-1,4-benzoxazine,thiomorpholinyl (also referred to as thiamorpholinyl),1,1-dioxothiomorpholinyl, piperidinyl, pyrrolidine, tetrahydrofuranyl,and the like.

Unless otherwise constrained by the definition for the heterocyclicsubstituent, such heterocyclic groups can be optionally substituted with1 to 5, or from 1 to 3 substituents, selected from alkoxy, substitutedalkoxy, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, acyl, acylamino, acyloxy, amino, substituted amino,aminoacyl, aminoacyloxy, oxyaminoacyl, azido, cyano, halogen, hydroxyl,oxo, thioketo, carboxyl, carboxylalkyl, thioaryloxy, thioheteroaryloxy,thioheterocyclooxy, thiol, thioalkoxy, substituted thioalkoxy, aryl,aryloxy, heteroaryl, heteroaryloxy, heterocyclyl, heterocyclooxy,hydroxyamino, alkoxyamino, nitro, —SO-alkyl, —SO-substituted alkyl,—SO-aryl, —SO-heteroaryl, —SO₂-alkyl, —SO₂-substituted alkyl, —SO₂-aryl,—SO₂-heteroaryl, and fused heterocycle.

“Heterocyclyloxy” refers to the group —O-heterocyclyl.

The term “heterocyclylthio” refers to the group heterocyclic-S—.

The term “heterocyclene” refers to the diradical group formed from aheterocycle, as defined herein.

The term “hydroxyamino” refers to the group —NHOH.

“Nitro” refers to the group —NO₂.

“Oxo” refers to the atom (═O).

“Sulfonyl” refers to the group SO₂-alkyl, SO₂-substituted alkyl,SO₂-alkenyl, SO₂-substituted alkenyl, SO₂-cycloalkyl, SO₂-substitutedcycloalkyl, SO₂-cycloalkenyl, SO₂-substituted cylcoalkenyl, SO₂-aryl,SO₂-substituted aryl, SO₂-heteroaryl, SO₂-substituted heteroaryl,SO₂-heterocyclic, and SO₂-substituted heterocyclic, wherein alkyl,substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substitutedalkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substitutedcycloalkenyl, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic are as definedherein. Sulfonyl includes, by way of example, methyl-SO₂—, phenyl-SO₂—,and 4-methylphenyl-SO₂—.

“Sulfonyloxy” refers to the group —OSO₂-alkyl, OSO₂-substituted alkyl,OSO₂-alkenyl, OSO₂-substituted alkenyl, OSO₂-cycloalkyl,OSO₂-substituted cycloalkyl, OSO₂-cycloalkenyl, OSO₂-substitutedcylcoalkenyl, OSO₂-aryl, OSO₂-substituted aryl, OSO₂-heteroaryl,OSO₂-substituted heteroaryl, OSO₂-heterocyclic, and OSO₂ substitutedheterocyclic, wherein alkyl, substituted alkyl, alkenyl, substitutedalkenyl, alkynyl, substituted alkynyl, cycloalkyl, substitutedcycloalkyl, cycloalkenyl, substituted cycloalkenyl, aryl, substitutedaryl, heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic are as defined herein.

The term “aminocarbonyloxy” refers to the group —OC(O)NRR where each Ris independently hydrogen, alkyl, substituted alkyl, aryl, heteroaryl,or heterocyclic wherein alkyl, substituted alkyl, aryl, heteroaryl andheterocyclic are as defined herein.

“Thiol” refers to the group —SH.

“Thioxo” or the term “thioketo” refers to the atom (═S).

“Alkylthio” or the term “thioalkoxy” refers to the group —S-alkyl,wherein alkyl is as defined herein. In certain embodiments, sulfur maybe oxidized to —S(O)—. The sulfoxide may exist as one or morestereoisomers.

The term “substituted thioalkoxy” refers to the group —S-substitutedalkyl.

The term “thioaryloxy” refers to the group aryl-S— wherein the arylgroup is as defined herein including optionally substituted aryl groupsalso defined herein.

The term “thioheteroaryloxy” refers to the group heteroaryl-S— whereinthe heteroaryl group is as defined herein including optionallysubstituted aryl groups as also defined herein.

The term “thioheterocyclooxy” refers to the group heterocyclyl-S—wherein the heterocyclyl group is as defined herein including optionallysubstituted heterocyclyl groups as also defined herein.

In addition to the disclosure herein, the term “substituted,” when usedto modify a specified group or radical, can also mean that one or morehydrogen atoms of the specified group or radical are each, independentlyof one another, replaced with the same or different substituent groupsas defined below.

In addition to the groups disclosed with respect to the individual termsherein, substituent groups for substituting for one or more hydrogens(any two hydrogens on a single carbon can be replaced with ═O, ═NR⁷⁰,═N—OR⁷⁰, ═N₂ or ═S) on saturated carbon atoms in the specified group orradical are, unless otherwise specified, —R⁶⁰, halo, ═O, —OR⁷⁰, —SR⁷⁰,—NR⁸⁰R⁸⁰, trihalomethyl, —CN, —OCN, —SCN, —NO, —NO₂, ═N₂, —N₃, —SO₂R⁷⁰,—SO₂O M⁺, —SO₂OR⁷⁰, —OSO₂R⁷⁰, —OSO₂O-M⁺, —OSO₂OR⁷⁰, —P(O)(O⁻)₂(M⁺)₂,—P(O)(OR⁷⁰)O⁻M⁺, —P(O)(OR⁷⁰)₂, —C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰,—C(O)O⁻M⁺, —C(O)OR⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰, —C(NR⁷⁰)NR⁸⁰R⁸⁰,—OC(O)R⁷⁰, —OC(S)R⁷⁰, —OC(O)O⁻M⁺, —OC(O)OR⁷⁰, —OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰,—NR⁷⁰C(S)R⁷⁰, —NR⁷⁰CO₂ ⁻M⁺, —NR⁷⁰CO₂R⁷⁰, —NR⁷⁰C(S)OR⁷⁰,—NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰ isselected from the group consisting of optionally substituted alkyl,cycloalkyl, heteroalkyl, heterocycloalkylalkyl, cycloalkylalkyl, aryl,arylalkyl, heteroaryl and heteroarylalkyl, each R⁷⁰ is independentlyhydrogen or R⁶⁰; each R⁸⁰ is independently R⁷⁰ or alternatively, twoR⁸⁰'s, taken together with the nitrogen atom to which they are bonded,form a 5-, 6- or 7-membered heterocycloalkyl which may optionallyinclude from 1 to 4 of the same or different additional heteroatomsselected from the group consisting of O, N and S, of which N may have —Hor C₁-C₃ alkyl substitution; and each M⁺ is a counter ion with a netsingle positive charge. Each M⁺ may independently be, for example, analkali ion, such as K⁺, Na⁺, Li⁺; an ammonium ion, such as ⁺N(R⁶⁰)₄; oran alkaline earth ion, such as [Ca²⁺]_(0.5), [Mg²⁺]_(0.5), or[Ba²⁺]_(0.5) (“subscript 0.5 means that one of the counter ions for suchdivalent alkali earth ions can be an ionized form of a compound of theinvention and the other a typical counter ion such as chloride, or twoionized compounds disclosed herein can serve as counter ions for suchdivalent alkali earth ions, or a doubly ionized compound of theinvention can serve as the counter ion for such divalent alkali earthions). As specific examples, —NR⁸⁰R⁸⁰ is meant to include —NH₂,—NH-alkyl, N-pyrrolidinyl, N-piperazinyl, 4N-methyl-piperazin-1-yl andN-morpholinyl.

In addition to the disclosure herein, substituent groups for hydrogenson unsaturated carbon atoms in “substituted” alkene, alkyne, aryl andheteroaryl groups are, unless otherwise specified, —R⁶⁰, halo, —O⁻M⁺,—OR⁷⁰, —SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰, trihalomethyl, —CF₃, —CN, —OCN, —SCN,—NO, —NO₂, —N₃, —SO₂R⁷⁰, —SO₃ ⁻M⁺, —SO₃R⁷⁰, —OSO₂R⁷⁰, —OSO₃ ⁻M⁺,—OSO₃R⁷⁰, —PO₃ ⁻²(M⁺)₂, —P(O)(OR⁷⁰)O⁻M⁺, —P(O)(OR⁷⁰)₂, —C(O)R⁷⁰,—C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰, —CO₂ ⁻M⁺, —CO₂R⁷⁰, —C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰,—C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰, —OCO₂ ⁻M⁺, —OCO₂R⁷⁰, —OC(S)OR⁷⁰,—NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰CO₂ ⁻M⁺, —NR⁷⁰CO₂R⁷⁰, —NR⁷⁰C(S)OR⁷⁰,—NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and —NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰,R⁷⁰, R⁸⁰ and M⁺ are as previously defined, provided that in case ofsubstituted alkene or alkyne, the substituents are not —O⁻M⁺, —OR⁷⁰,—SR⁷⁰, or —S⁻M⁺.

In addition to the groups disclosed with respect to the individual termsherein, substituent groups for hydrogens on nitrogen atoms in“substituted” heteroalkyl and cycloheteroalkyl groups are, unlessotherwise specified, —R⁶⁰, —O⁻M⁺, —OR⁷⁰, —SR⁷⁰, —S⁻M⁺, —NR⁸⁰R⁸⁰,trihalomethyl, —CF₃, —CN, —NO, —NO₂, —S(O)₂R⁷⁰, —S(O)₂O⁻M⁺, —S(O)₂OR⁷⁰,—OS(O)₂R⁷⁰, —OS(O)₂O⁻M⁺, —OS(O)₂OR⁷⁰, —P(O)(O⁻)₂(M⁺)₂, —P(O)(OR⁷⁰)O⁻M⁺,—P(O)(OR⁷⁰)(OR⁷⁰), —C(O)R⁷⁰, —C(S)R⁷⁰, —C(NR⁷⁰)R⁷⁰, —C(O)OR⁷⁰,—C(S)OR⁷⁰, —C(O)NR⁸⁰R⁸⁰, —C(NR⁷⁰)NR⁸⁰R⁸⁰, —OC(O)R⁷⁰, —OC(S)R⁷⁰,—OC(O)OR⁷⁰, —OC(S)OR⁷⁰, —NR⁷⁰C(O)R⁷⁰, —NR⁷⁰C(S)R⁷⁰, —NR⁷⁰C(O)OR⁷⁰,—NR⁷⁰C(S)OR⁷⁰, —NR⁷⁰C(O)NR⁸⁰R⁸⁰, —NR⁷⁰C(NR⁷⁰)R⁷⁰ and—NR⁷⁰C(NR⁷⁰)NR⁸⁰R⁸⁰, where R⁶⁰, R⁷⁰, R⁸⁰ and M⁺ are as previouslydefined.

In addition to the disclosure herein, in a certain embodiment, a groupthat is substituted has 1, 2, 3, or 4 substituents, 1, 2, or 3substituents, 1 or 2 substituents, or 1 substituent.

It is understood that in all substituted groups defined above, polymersarrived at by defining substituents with further substituents tothemselves (e.g., substituted aryl having a substituted aryl group as asubstituent which is itself substituted with a substituted aryl group,which is further substituted by a substituted aryl group, etc.) are notintended for inclusion herein. In such cases, the maximum number of suchsubstitutions is three. For example, serial substitutions of substitutedaryl groups specifically contemplated herein are limited to substitutedaryl-(substituted aryl)-substituted aryl.

Unless indicated otherwise, the nomenclature of substituents that arenot explicitly defined herein are arrived at by naming the terminalportion of the functionality followed by the adjacent functionalitytoward the point of attachment. For example, the substituent“arylalkyloxycarbonyl” refers to the group (aryl)-(alkyl)-O—C(O)—.

As to any of the groups disclosed herein which contain one or moresubstituents, it is understood, of course, that such groups do notcontain any substitution or substitution patterns which are stericallyimpractical and/or synthetically non-feasible. In addition, the subjectcompounds include all stereochemical isomers arising from thesubstitution of these compounds.

The term “pharmaceutically acceptable salt” means a salt which isacceptable for administration to a patient, such as a mammal (salts withcounterions having acceptable mammalian safety for a given dosageregime). Such salts can be derived from pharmaceutically acceptableinorganic or organic bases and from pharmaceutically acceptableinorganic or organic acids. “Pharmaceutically acceptable salt” refers topharmaceutically acceptable salts of a compound, which salts are derivedfrom a variety of organic and inorganic counter ions well known in theart and include, by way of example only, sodium, potassium, calcium,magnesium, ammonium, tetraalkylammonium, and the like; and when themolecule contains a basic functionality, salts of organic or inorganicacids, such as hydrochloride, hydrobromide, formate, tartrate, besylate,mesylate, acetate, maleate, oxalate, and the like.

The term “salt thereof” means a compound formed when a proton of an acidis replaced by a cation, such as a metal cation or an organic cation andthe like. Where applicable, the salt is a pharmaceutically acceptablesalt, although this is not required for salts of intermediate compoundsthat are not intended for administration to a patient. By way ofexample, salts of the present compounds include those wherein thecompound is protonated by an inorganic or organic acid to form a cation,with the conjugate base of the inorganic or organic acid as the anioniccomponent of the salt.

“Solvate” refers to a complex formed by combination of solvent moleculeswith molecules or ions of the solute. The solvent can be an organiccompound, an inorganic compound, or a mixture of both. Some examples ofsolvents include, but are not limited to, methanol,N,N-dimethylformamide, tetrahydrofuran, dimethylsulfoxide, and water.When the solvent is water, the solvate formed is a hydrate.

“Stereoisomer” and “stereoisomers” refer to compounds that have sameatomic connectivity but different atomic arrangement in space.Stereoisomers include cis-trans isomers, E and Z isomers, enantiomers,and diastereomers.

“Tautomer” refers to alternate forms of a molecule that differ only inelectronic bonding of atoms and/or in the position of a proton, such asenol-keto and imine-enamine tautomers, or the tautomeric forms ofheteroaryl groups containing a —N═C(H)—NH— ring atom arrangement, suchas pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles. Aperson of ordinary skill in the art would recognize that othertautomeric ring atom arrangements are possible.

It will be appreciated that the term “or a salt or solvate orstereoisomer thereof” is intended to include all permutations of salts,solvates and stereoisomers, such as a solvate of a pharmaceuticallyacceptable salt of a stereoisomer of subject compound.

“Pharmaceutically effective amount” and “therapeutically effectiveamount” refer to an amount of a compound sufficient to treat a specifieddisorder or disease or one or more of its symptoms and/or to prevent theoccurrence of the disease or disorder. In reference to tumorigenicproliferative disorders, a pharmaceutically or therapeutically effectiveamount comprises an amount sufficient to, among other things, cause thetumor to shrink or decrease the growth rate of the tumor.

By “treating” or “treatment” is meant that at least an amelioration ofthe symptoms associated with the condition afflicting the subject isachieved, where amelioration is used in a broad sense to refer to atleast a reduction in the magnitude of a parameter, e.g. symptom,associated with the condition being treated. As such, treatment alsoincludes situations where the pathological condition, or at leastsymptoms associated therewith, are completely inhibited, e.g., preventedfrom happening, or stopped, e.g. terminated, such that the subject nolonger suffers from the condition, or at least the symptoms thatcharacterize the condition. Thus treatment includes: (i) prevention,that is, reducing the risk of development of clinical symptoms,including causing the clinical symptoms not to develop, e.g., preventingdisease progression to a harmful state or prophylactic treatment of asubject; (ii) inhibition, that is, arresting the development or furtherdevelopment of clinical symptoms, e.g., mitigating or completelyinhibiting an active disease; and/or (iii) relief, that is, causing theregression of clinical symptoms or alleviating one or more symptoms ofthe disease or medical condition in the subject.

The terms “polypeptide,” “peptide,” and “protein” are usedinterchangeably herein to refer to a polymeric form of amino acids ofany length. Unless specifically indicated otherwise, “polypeptide,”“peptide,” and “protein” can include genetically coded and non-codedamino acids, chemically or biochemically modified or derivatized aminoacids, and polypeptides having modified peptide backbones. The termincludes fusion proteins, including, but not limited to, fusion proteinswith a heterologous amino acid sequence, fusions with heterologous andhomologous leader sequences, proteins which contain at least oneN-terminal methionine residue (e.g., to facilitate production in arecombinant host cell); immunologically tagged proteins; and the like.

“Native amino acid sequence” or “parent amino acid sequence” are usedinterchangeably herein to refer to the amino acid sequence of apolypeptide prior to modification to include a modified amino acidresidue.

The terms “amino acid analog,” “unnatural amino acid,” and the like maybe used interchangeably, and include amino acid-like compounds that aresimilar in structure and/or overall shape to one or more amino acidscommonly found in naturally occurring proteins (e.g., Ala or A, Cys orC, Asp or D, Glu or E, Phe or F, Gly or G, His or H, Ile or I, Lys or K,Leu or L, Met or M, Asn or N, Pro or P, Gln or Q, Arg or R, Ser or S,Thr or T, Val or V, Trp or W, Tyr or Y). Amino acid analogs also includenatural amino acids with modified side chains or backbones. Amino acidanalogs also include amino acid analogs with the same stereochemistry asin the naturally occurring D-form, as well as the L-form of amino acidanalogs. In some instances, the amino acid analogs share backbonestructures, and/or the side chain structures of one or more naturalamino acids, with difference(s) being one or more modified groups in themolecule. Such modification may include, but is not limited to,substitution of an atom (such as N) for a related atom (such as S),addition of a group (such as methyl, or hydroxyl, etc.) or an atom (suchas Cl or Br, etc.), deletion of a group, substitution of a covalent bond(single bond for double bond, etc.), or combinations thereof. Forexample, amino acid analogs may include α-hydroxy acids, and α-aminoacids, and the like.

The terms “amino acid side chain” or “side chain of an amino acid” andthe like may be used to refer to the substituent attached to theα-carbon of an amino acid residue, including natural amino acids,unnatural amino acids, and amino acid analogs. An amino acid side chaincan also include an amino acid side chain as described in the context ofthe modified amino acids and/or conjugates described herein.

As used herein the term “isolated” is meant to describe a compound ofinterest that is in an environment different from that in which thecompound naturally occurs. “Isolated” is meant to include compounds thatare within samples that are substantially enriched for the compound ofinterest and/or in which the compound of interest is partially orsubstantially purified.

As used herein, the term “substantially purified” refers to a compoundthat is removed from its natural environment and is at least 60% free,at least 75% free, at least 80% free, at least 85% free, at least 90%free, at least 95% free, at least 98% free, or more than 98% free, fromother components with which it is naturally associated.

The term “physiological conditions” is meant to encompass thoseconditions compatible with living cells, e.g., predominantly aqueousconditions of a temperature, pH, salinity, etc. that are compatible withliving cells.

As used herein, the term “amylin” refers to a 37 amino acid peptidehormone which is co-secreted with insulin from the pancreatic j-cell.

As used herein, the term “amyloid-beta protein” refers to peptides of36-43 amino acids resulting from cleavage of the amyloid precursorprotein, and which form the main component of neurotoxic amyloid plaquesfound in the brains of Alzheimer patients.

As used herein, the term “amylin receptor” refers to a receptor complexwhich binds amylin and amyloid-beta protein. The amylin receptorincludes the calcitonin receptor (CTR) dimerized with one of three knownsubtypes of receptor activity-modifying protein (RAMP1, RAMP2, RAMP3).Both amylin (HA) and amyloid-beta protein (Aβ42) bind and directlyactivate the amylin receptor and trigger biological and neurotoxiceffects. (Jhamandas et al., 2004).

As used herein, the term “amylin receptor antagonist” refers to acompound useful as an antagonist of the amylin receptor, or which bindsto, but does not activate, the amylin receptor. The amylin receptorantagonist displaces and blocks the binding of amylin or amyloid-betaprotein to the amylin receptor, thereby inhibiting the activity ofamylin or amyloid-beta protein.

As used herein, the term “AC253” refers to a peptide antagonist of theamylin receptor. The “AC” prefix indicates the peptide's identity withinthe peptide library of Amylin Pharmaceuticals Inc. As used herein, theterm “AC253” refers to a peptide having the amino acid sequence of SEQID NO: 1 (Ac-LGRLSQELHRLQTYPRTNTGSNTY) and which is capable of bindingto the amylin receptor, thereby inhibiting the activity of amylin,amyloid-beta protein, or both.

As used herein, the term “chronic administration” refers to repeatedadministration of a compound to a subject. In such treatment, thecompound can be administered at least once a week, such as at least oncea day, or at least twice or three times a day for a period of at leastone month, such as for example five months or more.

Before the present invention is further described, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may, of course, vary. It is also to be understoodthat the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to be limiting, sincethe scope of the present invention will be limited only by the appendedclaims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges, and are also encompassed within the invention, subjectto any specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination. All combinations of the embodimentspertaining to the invention are specifically embraced by the presentinvention and are disclosed herein just as if each and every combinationwas individually and explicitly disclosed, to the extent that suchcombinations embrace subject matter that are, for example, compoundsthat are stable compounds (i.e., compounds that can be made, isolated,characterized, and tested for biological activity). In addition, allsub-combinations of the various embodiments and elements thereof (e.g.,elements of the chemical groups listed in the embodiments describingsuch variables) are also specifically embraced by the present inventionand are disclosed herein just as if each and every such sub-combinationwas individually and explicitly disclosed herein.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology as “solely,” “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable sub-combination.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present invention isnot entitled to antedate such publication by virtue of prior invention.Further, the dates of publication provided may be different from theactual publication dates which may need to be independently confirmed.

DETAILED DESCRIPTION

The present disclosure provides compounds that are amylin receptorantagonists, compositions that include the subject compounds, andmethods for preparing and using the amylin receptor antagonists and thecompositions for treating, preventing, or ameliorating Alzheimer'sdisease.

Compounds and Methods of Treatment

The present disclosure provides methods of inhibiting activity of anamylin receptor. Embodiments of the present disclosure thus relate tomethods and uses of the compounds disclosed herein as amylin receptorantagonists which bind to, but do not activate, the amylin receptor.Compounds of the present disclosure may be used to displace and/or blockthe binding of amylin or amyloid-beta protein to the amylin receptor,thereby inhibiting the activity of amylin or amyloid-beta protein. Insome instances, compounds of the present disclosure are capable ofbinding to the AMY1 receptor. In some instances, compounds of thepresent disclosure are capable of binding to the AMY2 receptor. In someinstances, compounds of the present disclosure are capable of binding tothe AMY3 receptor. In some instances, compounds of the presentdisclosure are capable of binding to the AMY1 and AMY2 receptors. Insome instances, compounds of the present disclosure are capable ofbinding to the AMY1 and AMY3 receptors. In some instances, compounds ofthe present disclosure are capable of binding to the AMY2 and AMY3receptors. In some instances, compounds of the present disclosure arecapable of binding to the AMY1, AMY2 and AMY3 receptors. As used herein,“AMY1 receptor” refers to a heterodimeric complex of the calcitoninreceptor and RAMP1. As used herein, “AMY2 receptor” refers to aheterodimeric complex of the calcitonin receptor and RAMP2. As usedherein, “AMY3 receptor” refers to a heterodimeric complex of thecalcitonin receptor and RAMP3.

The amylin receptor antagonist may be used to reduce incidence of,reduce, treat, diminish, or prevent a disease or disorder in a subjectwhere it is of benefit to reduce amylin or amyloid-beta proteinactivity. In certain embodiments, the disease is Alzheimer's disease.Therapeutic uses of compounds of the present disclosure in diseases ordisorders, methods of prevention or treatment using compounds of thepresent disclosure, and uses of compounds of the present disclosure toprepare medicaments for therapeutic use are also included in embodimentsof the present disclosure. In certain instances, embodiments of thepresent disclosure relate to the therapeutic use of compounds of thepresent disclosure in humans.

In certain embodiments, a method of treating, preventing, orameliorating a disease or disorder in a subject is provided, where themethod includes administering to the subject a therapeutically effectiveamount of one or more compounds of the present disclosure or acomposition including same. As used herein, the term “disease” includes,but is not limited to, Alzheimer's disease. An effective amount of thecompound or composition may be an amount sufficient to provide eithersubjective relief of symptoms or an objectively identifiable improvementas noted by a clinician or other qualified observer. As such, methods of“treating”, “preventing” or “ameliorating” refer to interventionsperformed with the intention of alleviating the symptoms associatedwith, preventing the development of, or altering the pathology of adisease, disorder or condition, such as Alzheimer's disease. Thus, invarious embodiments, the methods of the present disclosure may includethe prevention (prophylaxis), moderation, reduction, or curing of adisease, disorder or condition at various stages, such as for exampleAlzheimer's disease. In various embodiments, therefore, those in need oftherapy/treatment may include those already having the disease, disorderor condition and/or those prone to, or at risk of developing, thedisease, disorder or condition and/or those in whom the disease,disorder or condition is to be prevented.

In certain embodiments, the amylin receptor antagonist of the presentdisclosure is effective for reducing cyclic AMP (cAMP) signal productionin a cell. For example, administration of a therapeutically effectiveamount of the amylin receptor antagonist may cause a reduction in cAMPsignal production in a cell as compared to a cell that has not beenadministered the amylin receptor antagonist.

In certain embodiments, compounds of the present disclosure produce aneuroprotective effect against amylin and/or amyloid-beta proteininduced neurotoxicity. For example, in some cases, administration of acompound of the present disclosure is therapeutically effective forprotecting neurons against the neurotoxic effect of amyloid-betaprotein. In some cases, administration of a compound of the presentdisclosure is therapeutically effective for protecting neurons againstthe neurotoxic effect of amylin.

Methods of the present disclosure include administering to a subject inneed thereof, a therapeutically effective amount of an amylin receptorantagonist. In certain embodiments, the amylin receptor antagonist is anon-peptidic compound. Non-peptidic compounds according to the presentdisclosure do not contain as part of their chemical structure a peptideor peptide derivative (e.g., modified peptide).

Formula (I)

In certain embodiments, the amylin receptor antagonist is a compound offormula (I):

wherein:

R is selected from the group consisting of —H, C₁-C₆-alkyl, andsubstituted C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H,C₁-C₆-alkyl, and substituted C₁-C₆-alkyl;

each W is selected from the group consisting of —CH₂—, —CHR³— and—CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, substituted carbocycle or oxo (i.e., ═O);

m is selected from 1, 2 or 3;

Q is selected from the group consisting of —CH₂—, —C(═O)—, —CH₂C(═O)—,and —CH₂CH₂—;

X is present or absent, and if present is selected from the groupconsisting of —CH₂— and —C(═O)—;

Y is selected from the group consisting of —CH₂—, and —C(═O)—;

Z is selected from the group consisting of —OR⁵, halogen, —CN, —NR⁵R⁵,—NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and

each R⁵ is independently selected from the group consisting of —H,C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, R is selected from —H, C₁-C₆-alkyl, andsubstituted C₁-C₆-alkyl. For example, in some embodiments, R can be —H.In some embodiments, R can be C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl).

In certain embodiments, each R¹ is independently selected from the groupconsisting of H, halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. For example, R¹ can be H, halogen, C₁-C₆-alkyl,substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,—OR², heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, or substituted heteroaryl. In some instances, R¹ is H. Insome instances, R¹ is halogen (e.g., F, Cl, Br, I). In some instances,R¹ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl)or substituted C₁-C₆-alkyl (e.g., substituted methyl (e.g., —CF₃),substituted ethyl, substituted propyl, substituted butyl, substitutedpentyl, or substituted hexyl). In some instances, R¹ is C₃-C₆-cycloalkyl(e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) orsubstituted C₃-C₆-cycloalkyl (e.g., substituted cyclopropyl, substitutedcyclobutyl, substituted cyclopentyl, or substituted cyclohexyl). In someinstances, R¹ is —OR². In some instances, R¹ is heterocyclyl orsubstituted heterocyclyl (e.g., unsubstituted or substitutedpyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, andthe like). In some instances, R¹ is aryl or substituted aryl (e.g.,unsubstituted or substituted phenyl). In some instances, R¹ isheteroaryl or substituted heteroaryl (e.g., unsubstituted or substitutedpyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, and the like).

In certain embodiments, each R² is independently selected from the groupconsisting of —H, C₁-C₆-alkyl, and substituted C₁-C₆-alkyl. For example,in some embodiments, R² can be —H. In some embodiments, R² can beC₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl) orsubstituted C₁-C₆-alkyl (e.g., substituted methyl, substituted ethyl,substituted propyl, substituted butyl, substituted pentyl, orsubstituted hexyl).

In certain embodiments, each W is selected from the group consisting of—CH₂—, —CHR³— and —CR³R⁴—. In some instances, W is —CH₂—. In someinstances, W is —CHR³—. In some instances, W is —CR³R⁴—.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, and substitutedfused-heterocycle, or together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo.

For example, R³ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R³ is halogen (e.g., F, Cl, Br,I). In some instances, R³ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R³ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R³ is heterocyclyl orsubstituted heterocyclyl. In some instances, R³ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R³is heteroaryl or substituted heteroaryl. In some instances, R³ isfused-heterocycle or substituted fused-heterocycle.

For example, R⁴ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R⁴ is halogen (e.g., F, Cl, Br,I). In some instances, R⁴ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R⁴ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R⁴ is heterocyclyl orsubstituted heterocyclyl. In some instances, R⁴ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R⁴is heteroaryl or substituted heteroaryl. In some instances, R⁴ isfused-heterocycle or substituted fused-heterocycle.

In certain embodiments, together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo (i.e., ═O). In some instances, together R³and R⁴ comprise a carbocycle, such as a C₃-C₆ carbocycle (e.g.,cyclopropyl, cyclobutyl, cyclopentyl). In some instances, together R³and R⁴ comprise a substituted carbocycle, such as a substituted C₃-C₆carbocycle (e.g., substituted cyclopropyl, substituted cyclobutyl,substituted cyclopentyl). In some instances, together R³ and R⁴ comprisean oxo group (i.e., ═O).

In certain embodiments, m is selected from 1, 2 or 3. In some instances,m is 1. In some instances, m is 2. In some instances, m is 3.

In certain embodiments, Q is selected from —CH₂—, —C(═O)—, —CH₂C(═O)—,and —CH₂CH₂—. In some instances, Q is —CH₂—. In some instances, Q is—C(═O)—. In some instances, Q is —CH₂C(═O)—. In some instances, Q is—CH₂CH₂—.

In certain embodiments, X is present or absent, and if present isselected from the group consisting of —CH₂— and —C(═O)—. In someinstances, X is absent. In embodiments were X is absent, the N adjacentto X is directly bonded to the phenyl ring in formula (I). In someinstances, X is present. In some instances, X is —CH₂—. In someinstances, X is —C(═O)—.

In certain embodiments, Y is selected from —CH₂—, and —C(═O)—. In someinstances, Y is —CH₂—. In some instances, Y is —C(═O)—.

In certain embodiments, Z is selected from the group consisting of —OR⁵,halogen, —CN, —NR⁵R⁵, —NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl.In some instances, Z is —OR⁵. In some instances, Z is halogen (e.g., F,Cl, Br, I). In some instances, Z is —CN. In some instances, Z is —NR⁵R⁵.In some instances, Z is —NHC(═O)R⁵. In some instances, Z is—NHC(═O)NR⁵R⁵. In some instances, Z is aryl (e.g., phenyl). In someinstances, Z is heteroaryl.

In certain embodiments, each R⁵ is independently selected from the groupconsisting of —H, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl,substituted aryl, heteroaryl, and substituted heteroaryl. For example,R⁵ can be H, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substitutedaryl, heteroaryl, or substituted heteroaryl. In some instances, R⁵ is H.In some instances, R⁵ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl (e.g., isopropyl), substituted propyl,substituted butyl, substituted pentyl, or substituted hexyl). In someinstances, R⁵ is aryl or substituted aryl (e.g., unsubstituted orsubstituted phenyl). In some instances, R⁵ is heteroaryl or substitutedheteroaryl (e.g., unsubstituted or substituted pyrrolyl, pyrazolyl,imidazolyl, pyridinyl, pyrimidinyl, and the like).

Formula (II)

In certain embodiments, the amylin receptor antagonist is a compound offormula (II):

wherein:

R is selected from the group consisting of —H and C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H, andC₁-C₆-alkyl;

each W is selected from the group consisting of —CH₂—, —CHR³— and—CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, substituted carbocycle or oxo;

m is selected from 1, 2 or 3;

Q is selected from the group consisting of —CH₂—, —C(═O)— and—CH₂C(═O)—;

Y is selected from the group consisting of —CH₂—, and —C(═O)—;

Z is selected from the group consisting of —OR⁵, -halogen, —NR⁵R⁵,—NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and

each R⁵ is independently selected from the group consisting of —H,C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, R is selected from —H and C₁-C₆-alkyl. Forexample, in some embodiments, R can be —H. In some embodiments, R can beC₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl).

In certain embodiments, each R¹ is independently selected from the groupconsisting of H, halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. For example, R¹ can be H, halogen, C₁-C₆-alkyl,substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,—OR², heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, or substituted heteroaryl. In some instances, R¹ is H. Insome instances, R¹ is halogen (e.g., F, Cl, Br, I). In some instances,R¹ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl)or substituted C₁-C₆-alkyl (e.g., substituted methyl (e.g., —CF₃),substituted ethyl, substituted propyl, substituted butyl, substitutedpentyl, or substituted hexyl). In some instances, R¹ is C₃-C₆-cycloalkyl(e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) orsubstituted C₃-C₆-cycloalkyl (e.g., substituted cyclopropyl, substitutedcyclobutyl, substituted cyclopentyl, or substituted cyclohexyl). In someinstances, R¹ is —OR². In some instances, R¹ is heterocyclyl orsubstituted heterocyclyl (e.g., unsubstituted or substitutedpyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, andthe like). In some instances, R¹ is aryl or substituted aryl (e.g.,unsubstituted or substituted phenyl). In some instances, R¹ isheteroaryl or substituted heteroaryl (e.g., unsubstituted or substitutedpyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, and the like).

In certain embodiments, each R² is independently selected from the groupconsisting of —H and C₁-C₆-alkyl. For example, in some embodiments, R²can be —H. In some embodiments, R² can be C₁-C₆-alkyl (e.g., methyl,ethyl, propyl, butyl, pentyl, or hexyl).

In certain embodiments, each W is selected from the group consisting of—CH₂—, —CHR³— and —CR³R⁴—. In some instances, W is —CH₂—. In someinstances, W is —CHR³—. In some instances, W is —CR³R⁴—.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, and substitutedfused-heterocycle, or together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo.

For example, R³ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R³ is halogen (e.g., F, Cl, Br,I). In some instances, R³ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R³ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R³ is heterocyclyl orsubstituted heterocyclyl. In some instances, R³ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R³is heteroaryl or substituted heteroaryl. In some instances, R³ isfused-heterocycle or substituted fused-heterocycle.

For example, R⁴ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R⁴ is halogen (e.g., F, Cl, Br,I). In some instances, R⁴ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R⁴ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R⁴ is heterocyclyl orsubstituted heterocyclyl. In some instances, R⁴ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R⁴is heteroaryl or substituted heteroaryl. In some instances, R⁴ isfused-heterocycle or substituted fused-heterocycle.

In certain embodiments, together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo (i.e., ═O). In some instances, together R³and R⁴ comprise a carbocycle, such as a C₃-C₆ carbocycle (e.g.,cyclopropyl, cyclobutyl, cyclopentyl). In some instances, together R³and R⁴ comprise a substituted carbocycle, such as a substituted C₃-C₆carbocycle (e.g., substituted cyclopropyl, substituted cyclobutyl,substituted cyclopentyl). In some instances, together R³ and R⁴ comprisean oxo group (i.e., ═O).

In certain embodiments, m is selected from 1, 2 or 3. In some instances,m is 1. In some instances, m is 2. In some instances, m is 3.

In certain embodiments, Q is selected from —CH₂—, —C(═O)—, and—CH₂C(═O)—. In some instances, Q is —CH₂—. In some instances, Q is—C(═O)—. In some instances, Q is —CH₂C(═O)—.

In certain embodiments, Y is selected from —CH₂—, and —C(═O)—. In someinstances, Y is —CH₂—. In some instances, Y is —C(═O)—.

In certain embodiments, Z is selected from the group consisting of —OR⁵,halogen, —NR⁵R⁵, —NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl. Insome instances, Z is —OR⁵. In some instances, Z is halogen (e.g., F, Cl,Br, I). In some instances, Z is —NR⁵R⁵. In some instances, Z is—NHC(═O)R⁵. In some instances, Z is —NHC(═O)NR⁵R⁵. In some instances, Zis aryl (e.g., phenyl). In some instances, Z is heteroaryl.

In certain embodiments, each R⁵ is independently selected from the groupconsisting of —H, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl,substituted aryl, heteroaryl, and substituted heteroaryl. For example,R⁵ can be H, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substitutedaryl, heteroaryl, or substituted heteroaryl. In some instances, R⁵ is H.In some instances, R⁵ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl (e.g., isopropyl), substituted propyl,substituted butyl, substituted pentyl, or substituted hexyl). In someinstances, R⁵ is aryl or substituted aryl (e.g., unsubstituted orsubstituted phenyl). In some instances, R⁵ is heteroaryl or substitutedheteroaryl (e.g., unsubstituted or substituted pyrrolyl, pyrazolyl,imidazolyl, pyridinyl, pyrimidinyl, and the like).

Formula (III)

In certain embodiments, the amylin receptor antagonist is a compound offormula (III):

wherein:

R is selected from the group consisting of —H and C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H, andC₁-C₆-alkyl;

each W is selected from the group consisting of —CH₂—, —CHR³— and—CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, substituted carbocycle or oxo;

m is selected from 1, 2 or 3;

Z is selected from the group consisting of —OR⁵, halogen, —NR⁵R⁵,—NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and

each R⁵ is independently selected from the group consisting of —H,C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, R is selected from —H and C₁-C₆-alkyl. Forexample, in some embodiments, R can be —H. In some embodiments, R can beC₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl).

In certain embodiments, each R¹ is independently selected from the groupconsisting of H, halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. For example, R¹ can be H, halogen, C₁-C₆-alkyl,substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,—OR², heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, or substituted heteroaryl. In some instances, R¹ is H. Insome instances, R¹ is halogen (e.g., F, Cl, Br, I). In some instances,R¹ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl)or substituted C₁-C₆-alkyl (e.g., substituted methyl (e.g., —CF₃),substituted ethyl, substituted propyl, substituted butyl, substitutedpentyl, or substituted hexyl). In some instances, R¹ is C₃-C₆-cycloalkyl(e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) orsubstituted C₃-C₆-cycloalkyl (e.g., substituted cyclopropyl, substitutedcyclobutyl, substituted cyclopentyl, or substituted cyclohexyl). In someinstances, R¹ is —OR². In some instances, R¹ is heterocyclyl orsubstituted heterocyclyl (e.g., unsubstituted or substitutedpyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl, morpholinyl, andthe like). In some instances, R¹ is aryl or substituted aryl (e.g.,unsubstituted or substituted phenyl). In some instances, R¹ isheteroaryl or substituted heteroaryl (e.g., unsubstituted or substitutedpyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, and the like).

In certain embodiments, each R² is independently selected from the groupconsisting of —H and C₁-C₆-alkyl. For example, in some embodiments, R²can be —H. In some embodiments, R² can be C₁-C₆-alkyl (e.g., methyl,ethyl, propyl, butyl, pentyl, or hexyl).

In certain embodiments, each W is selected from the group consisting of—CH₂—, —CHR³— and —CR³R⁴—. In some instances, W is —CH₂—. In someinstances, W is —CHR³—. In some instances, W is —CR³R⁴—.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, and substitutedfused-heterocycle, or together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo.

For example, R³ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R³ is halogen (e.g., F, Cl, Br,I). In some instances, R³ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R³ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R³ is heterocyclyl orsubstituted heterocyclyl. In some instances, R³ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R³is heteroaryl or substituted heteroaryl. In some instances, R³ isfused-heterocycle or substituted fused-heterocycle.

For example, R⁴ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R⁴ is halogen (e.g., F, Cl, Br,I). In some instances, R⁴ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R⁴ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R⁴ is heterocyclyl orsubstituted heterocyclyl. In some instances, R⁴ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R⁴is heteroaryl or substituted heteroaryl. In some instances, R⁴ isfused-heterocycle or substituted fused-heterocycle.

In certain embodiments, together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo (i.e., ═O). In some instances, together R³and R⁴ comprise a carbocycle, such as a C₃-C₆ carbocycle (e.g.,cyclopropyl, cyclobutyl, cyclopentyl). In some instances, together R³and R⁴ comprise a substituted carbocycle, such as a substituted C₃-C₆carbocycle (e.g., substituted cyclopropyl, substituted cyclobutyl,substituted cyclopentyl). In some instances, together R³ and R⁴ comprisean oxo group (i.e., ═O).

In certain embodiments, m is selected from 1, 2 or 3. In some instances,m is 1. In some instances, m is 2. In some instances, m is 3.

In certain embodiments, Z is selected from the group consisting of —OR⁵,halogen, —NR⁵R⁵, —NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl. Insome instances, Z is —OR⁵. In some instances, Z is halogen (e.g., F, Cl,Br, I). In some instances, Z is —NR⁵R⁵. In some instances, Z is—NHC(═O)R⁵. In some instances, Z is —NHC(═O)NR⁵R⁵. In some instances, Zis aryl (e.g., phenyl). In some instances, Z is heteroaryl.

In certain embodiments, each R⁵ is independently selected from the groupconsisting of —H, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl, andheteroaryl. For example, R⁵ can be H, C₁-C₆-alkyl, substitutedC₁-C₆-alkyl, aryl, substituted aryl, heteroaryl, or substitutedheteroaryl. In some instances, R⁵ is H. In some instances, R⁵ isC₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl) orsubstituted C₁-C₆-alkyl (e.g., substituted methyl, substituted ethyl(e.g., isopropyl), substituted propyl, substituted butyl, substitutedpentyl, or substituted hexyl). In some instances, R⁵ is aryl (e.g.,phenyl). In some instances, R⁵ is heteroaryl (e.g., pyrrolyl, pyrazolyl,imidazolyl, pyridinyl, pyrimidinyl, and the like).

Compounds of the present disclosure (e.g., compounds of formulae (I),(II) and (III) as described herein) also include an enantiomer, amixture of enantiomers, a mixture of two or more diastereomers, atautomer, a mixture of two or more tautomers, or an isotopic variantthereof.

In addition, compounds of the present disclosure (e.g., compounds offormulae (I), (II) and (III) as described herein) also include apharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, compounds of the present disclosure (e.g.,compounds that find use in the methods of the present disclosure)include compounds selected from:

1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione;

1-(3-methoxypropyl)-4-methylquinoxaline-2,3(1H,4H)-dione;

1-(3-hydroxypropyl)-4-methylquinoxaline-2,3(1H,4H)-dione;

1-(3-methoxypropyl)quinoxaline-2,3(1H,4H)-dione;

1-(3-phenoxypropyl)quinoxaline-2,3(1H,4H)-dione;

1-(2-hydroxyethyl)quinoxaline-2,3(1H,4H)-dione;

1-(3-hydroxypropyl)-1H-benzo[b][1,4]diazepine-2,4(3H,5H)-dione;

1-(3-hydroxypropyl)-6-(pyridin-3-yl)quinoxaline-2,3(1H,4H)-dione;

1-(3-hydroxypropyl)-6-methylquinoxaline-2,3(1H,4H)-dione;

1-(3-hydroxypropyl)-7-(pyridin-3-yl)quinoxaline-2,3(1H,4H)-dione;

1-(3-hydroxypropyl)-6,7-dimethylquinoxaline-2,3(1H,4H)-dione;

5-fluoro-1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione;

5-chloro-1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione;

6,7-dichloro-1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione;

6-chloro-1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione;

1-(3-hydroxypropyl)-6-(trifluoromethyl)quinoxaline-2,3(1H,4H)-dione;

1-(3-hydroxypropyl)-7-(trifluoromethyl)quinoxaline-2,3(1H,4H)-dione;

7-chloro-6-fluoro-1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione;

4-(3,3,3-trifluoropropanoyl)-3,4-dihydroquinoxalin-2(1H)-one;

3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propanenitrile;

N-(3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl)acetamide;

4-(4-hydroxybutanoyl)-3,4-dihydroquinoxalin-2(1H)-one; and

4-oxo-4-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)butanenitrile.

Formula (IV)

In certain embodiments, the amylin receptor antagonist is a compound offormula (IV):

wherein:

R is selected from the group consisting of —H and C₁-C₆-alkyl;

each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl;

each R² is independently selected from the group consisting of —H, andC₁-C₆-alkyl;

each W is independently selected from the group consisting of —CH₂—,—CHR³— and —CR³R⁴—;

R³ and R⁴ are independently selected from the group consisting ofhalogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl,aryl, substituted aryl, heteroaryl, substituted heteroaryl,fused-heterocycle, and substituted fused-heterocycle, or together R³ andR⁴ comprise a carbocycle, or substituted carbocycle;

Z is selected from the group consisting of —OH, —OC(═O)CH₃, —OC(═O)Ph,and —NHC(═O)CH₃;

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof;

with the proviso that the compound is not:

N-[3-(3,4-dihydro-3-oxo-1(2H)-quinoxalinyl)-3-oxopropyl]acetamide.

In certain embodiments, R is selected from —H and C₁-C₆-alkyl. Forexample, in some embodiments, R can be —H. In some embodiments, R can beC₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl).

In certain embodiments, each R¹ is independently selected from the groupconsisting of H, halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl. For example, R¹ can be H, halogen, C₁-C₆-alkyl,substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,—OR², heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, or substituted heteroaryl. In some instances, R¹ is H. Insome instances, R¹ is halogen (e.g., F, Cl, Br, I). In some instances,R¹ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl)or substituted C₁-C₆-alkyl (e.g., substituted methyl (e.g., —CF₃),substituted ethyl, substituted propyl, substituted butyl, substitutedpentyl, or substituted hexyl). In some instances, R¹ is C₃-C₆-cycloalkyl(e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) orsubstituted C₃-C₆-cycloalkyl (e.g., substituted cyclopropyl, substitutedcyclobutyl, substituted cyclopentyl, or substituted cyclohexyl). In someinstances, R¹ is —OR², such as —OCH₃. In some instances, R¹ isheterocyclyl or substituted heterocyclyl (e.g., unsubstituted orsubstituted pyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl,morpholinyl, and the like). In some instances, R¹ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R¹is heteroaryl or substituted heteroaryl (e.g., unsubstituted orsubstituted pyrrolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, andthe like).

In certain embodiments, each R² is independently selected from the groupconsisting of —H and C₁-C₆-alkyl. For example, in some embodiments, R²can be —H. In some embodiments, R² can be C₁-C₆-alkyl (e.g., methyl,ethyl, propyl, butyl, pentyl, or hexyl).

In certain embodiments, each W is selected from the group consisting of—CH₂—, —CHR³— and —CR³R⁴—. In some instances, W is —CH₂—. In someinstances, W is —CHR³—. In some instances, W is —CR³R⁴—.

In certain embodiments, R³ and R⁴ are independently selected from thegroup consisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, and substitutedfused-heterocycle, or together R³ and R⁴ comprise a carbocycle orsubstituted carbocycle.

For example, R³ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R³ is halogen (e.g., F, Cl, Br,I). In some instances, R³ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R³ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R³ is heterocyclyl orsubstituted heterocyclyl. In some instances, R³ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R³is heteroaryl or substituted heteroaryl. In some instances, R³ isfused-heterocycle or substituted fused-heterocycle.

For example, R⁴ can be halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, or substitutedfused-heterocycle. In some instances, R⁴ is halogen (e.g., F, Cl, Br,I). In some instances, R⁴ is C₁-C₆-alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl) or substituted C₁-C₆-alkyl (e.g., substitutedmethyl, substituted ethyl, substituted propyl, substituted butyl,substituted pentyl, or substituted hexyl). In some instances, R⁴ isC₃-C₆-cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl) or substituted C₃-C₆-cycloalkyl (e.g., substitutedcyclopropyl, substituted cyclobutyl, substituted cyclopentyl, orsubstituted cyclohexyl). In some instances, R⁴ is heterocyclyl orsubstituted heterocyclyl. In some instances, R⁴ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R⁴is heteroaryl or substituted heteroaryl. In some instances, R⁴ isfused-heterocycle or substituted fused-heterocycle.

In certain embodiments, together R³ and R⁴ comprise a carbocycle orsubstituted carbocycle. In some instances, together R³ and R⁴ comprise acarbocycle, such as a C₃-C₆ carbocycle (e.g., cyclopropyl, cyclobutyl,cyclopentyl). In some instances, together R³ and R⁴ comprise asubstituted carbocycle, such as a substituted C₃-C₆ carbocycle (e.g.,substituted cyclopropyl, substituted cyclobutyl, substitutedcyclopentyl.

In certain embodiments, Z is selected from the group consisting of —OH,—OC(═O)CH₃, —OC(═O)Ph, and —NHC(═O)CH₃. In some instances, Z is —OH. Insome instances, Z is —OC(═O)CH₃. In some instances, Z is —OC(═O)Ph. Insome instances, Z is —NHC(═O)CH₃.

Compounds of the present disclosure (e.g., compounds of formula (IV) asdescribed herein) also include an enantiomer, a mixture of enantiomers,a mixture of two or more diastereomers, a tautomer, a mixture of two ormore tautomers, or an isotopic variant thereof.

In addition, compounds of the present disclosure (e.g., compounds offormula (IV) as described herein) also include a pharmaceuticallyacceptable salt, solvate, or hydrate thereof.

In certain embodiments, compounds of the present disclosure (e.g.,compounds that find use in the methods of the present disclosure)include compounds selected from:

3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl acetate;

4-(3-(2-hydroxyethylamino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one;

4-(3-(dimethylamino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one;

4-(3-hydroxypropanoyl)-3,4-dihydroquinoxalin-2(1H)-one;

N,N-dimethyl-3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propanamide;

4-(3-hydroxypropanoyl)-4,5-dihydro-1H-benzo[e][1,4]diazepin-2(3H)-one;

3-hydroxy-1-(4-methoxy-2,3-dihydro-1H-benzo[b][1,4]diazepin-1-yl)propan-1-one;

1-(3-hydroxypropanoyl)-3,4-dihydro-1H-benzo[e][1,4]diazepin-5(2H)-one;

5-(3-hydroxypropanoyl)-4,5-dihydro-1H-benzo[b][1,4]diazepin-2(3H)-one;

6,7-difluoro-4-(3-hydroxypropanoyl)-3,4-dihydroquinoxalin-2(1H)-one;

4-(3-hydroxybutanoyl)-3,4-dihydroquinoxalin-2(1H)-one;

4-(3-hydroxypropanoyl)-6,7-dimethyl-3,4-dihydroquinoxalin-2(1H)-one;

4-(3-hydroxycyclobutanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one;

2-hydroxy-N-(3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl)acetamide;

4-(3-(7-nitrobenzo[c][1,2,5]oxadiazol-4-ylamino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one;and

4-(2-hydroxycyclopentanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one.

In certain embodiments, the compound of formula (IV) does not includeN-[3-(3,4-dihydro-3-oxo-1(2H)-quinoxalinyl)-3-oxopropyl]acetamide.

Formula (V), Formula (VI) and Formula (VII)

In certain embodiments, the amylin receptor antagonist is a compound offormula (V), formula (VI) or formula (VII):

wherein:

R⁷ is selected from the group consisting of —CH₂OC(═O)OR⁸,—C(CH₃)HOC(═O)OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NHR⁸, —C(═O)NR⁸R⁸, andC₁-C₆-alkyl; and

each R⁸ is independently selected from C₁-C₆-alkyl, substitutedC₂-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,heterocycyl, substituted heterocycyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl,

or an enantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof;

or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, R⁷ is selected from the group consisting of—CH₂OC(═O)OR⁸, —C(CH₃)HOC(═O)OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NHR⁸,—C(═O)NR⁸R⁸, and C₁-C₆-alkyl. In some instances, R⁷ is —CH₂OC(═O)OR⁸. Insome instances, R⁷ is —C(CH₃)HOC(═O)OR⁸. In some instances, R⁷ is—C(═O)R⁸. In some instances, R⁷ is —C(═O)OR⁸. In some instances, R⁷ is—C(═O)NHR⁸. In some instances, R⁷ is —C(═O)NR⁸R⁸. In some instances, R⁷is C₁-C₆-alkyl.

In certain embodiments, each R⁸ is independently selected fromC₁-C₆-alkyl, substituted C₂-C₆-alkyl, C₃-C₆-cycloalkyl, substitutedC₃-C₆-cycloalkyl, heterocycyl, substituted heterocycyl, aryl,substituted aryl, heteroaryl, and substituted heteroaryl. For example,R⁸ can be C₁-C₆-alkyl alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl,or hexyl) or substituted C₂-C₆-alkyl (e.g., substituted ethyl,substituted propyl, substituted butyl, substituted pentyl, orsubstituted hexyl). In some instances, R⁸ is C₃-C₆-cycloalkyl (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) or substitutedC₃-C₆-cycloalkyl (e.g., substituted cyclopropyl, substituted cyclobutyl,substituted cyclopentyl, or substituted cyclohexyl). In some instances,R⁸ is heterocyclyl or substituted heterocyclyl (e.g., unsubstituted orsubstituted pyrrolidinyl, imidazolidinyl, piperidinyl, piperazinyl,morpholinyl, and the like). In some instances, R⁸ is aryl or substitutedaryl (e.g., unsubstituted or substituted phenyl). In some instances, R⁸is heteroaryl or substituted heteroaryl (e.g., unsubstituted orsubstituted pyrrolyl, pyrazolyl, imidazolyl, pyridinyl, pyrimidinyl, andthe like).

Compounds of the present disclosure (e.g., compounds of formula (V),formula (VI) or formula (VII) as described herein) also include anenantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof.

In addition, compounds of the present disclosure (e.g., compounds offormula (V), formula (VI) or formula (VII) as described herein) alsoinclude a pharmaceutically acceptable salt, solvate, or hydrate thereof.

In certain embodiments, compounds of the present disclosure (e.g.,compounds that find use in the methods of the present disclosure)include compounds selected from:

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl acetate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl isobutyrate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl2-amino-3-methylbutanoate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl benzoate;

Compound 46

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl 2-methylbenzoate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl 4-methylbenzoate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl 2-methoxybenzoate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl ethyl carbonate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl isopropyl carbonate;

(3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propoxy)methyl ethylcarbonate;

1-(3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propoxy)ethyl ethylcarbonate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl cyclopentylcarbamate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl cyclohexylcarbamate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl dimethylcarbamate;

3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propylpyrrolidine-1-carboxylate;

isopropyl4-(3-hydroxypropyl)-2,3-dioxo-3,4-dihydroquinoxaline-1(2H)-carboxylate;

(4-(3-hydroxypropyl)-2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)methylmethyl carbonate;

ethyl(4-(3-hydroxypropyl)-2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)methylcarbonate;

4-(3-hydroxypropyl)-3-oxo-3,4-dihydroquinoxalin-2-yl isopropylcarbonate; and

(4-(3-hydroxypropyl)-3-oxo-3,4-dihydroquinoxalin-2-yloxy)methyl methylcarbonate.

In certain embodiments, a compound of formula (V), formula (VI) andformula (VII) is a prodrug of a compound of formula (I), formula (II) orformula (III) as described herein. Embodiments of prodrugs of compoundsof formula (IV) are also provided by the disclosure herein.

The term “prodrug” refers to a derivative of an active compound (e.g., adrug or active agent) that undergoes a transformation under theconditions of use, such as within the body, to release the activecompound. Prodrugs may be, but are not necessarily, pharmacologicallyinactive until converted into the active drug. In some cases, prodrugsare pharmacologically inactive until converted into the active drug. Incertain cases, compounds that include a progroup may facilitate removalof the progroup at a desired site of action for the pharmaceuticallyactive form of the compound, or after a desired amount of time afteradministration of the compound (e.g., delayed release formulations,controlled release formulations, and the like). A wide variety ofprogroups, as well as the resultant promoieties, suitable for maskingfunctional groups in the active drugs to yield prodrugs may be used. Forexample, a hydroxyl functional group can be masked as a sulfonate, esteror carbonate promoiety, which can be hydrolyzed in vivo to provide thehydroxyl group. An amino functional group can be masked as an amide,carbamate, imine, urea, phosphenyl, phosphoryl or sulfenyl promoiety,which can be hydrolyzed in vivo to provide the amino group. A carboxylgroup can be masked as an ester (including silyl esters and thioesters),amide or hydrazide promoiety, which can be hydrolyzed in vivo to providethe carboxyl group. Other specific examples of suitable progroups andtheir respective promoieties will be apparent to those of skill in theart.

In some instances, prodrugs may be obtained by masking a functionalgroup in the drug believed to be in part required for activity with aprogroup to form a promoiety which undergoes a transformation, such ascleavage, under the specified conditions of use to release the activedrug. The cleavage of the promoiety can proceed spontaneously, such asby way of a hydrolysis or oxidation reaction, or it can be catalyzed orinduced by another agent, such as by an enzyme (e.g., cytochrome P450,an esterase, a peptidase, and the like), by light, by acid, or by achange of or exposure to a physical or environmental parameter, such asa change of temperature. The agent can be endogenous to the conditionsof use, such as an enzyme present in the cells to which the prodrug isadministered or the acidic conditions of the stomach, or it can besupplied exogenously. For example, in certain cases, compounds thatinclude a progroup may facilitate an increase in gastrointestinalpermeability, an increase in gastrointestinal absorption, and/or anincrease in solubility of the compound.

The prodrugs of the present disclosure include compounds of the formulaedescribed herein. Pharmaceutical compositions of the prodrugs andmethods of use involving the prodrugs of the present disclosure are alsocontemplated herein.

Methods of Use

The compounds of the present disclosure find use in treatment of acondition or disease in a subject that is amenable to treatment byadministration of the compound. Thus, in some embodiments, provided aremethods that include administering to a subject a therapeuticallyeffective amount of any of the compounds of the present disclosure. Incertain aspects, provided are methods of delivering a compound to atarget site in a subject, the method including administering to thesubject a pharmaceutical composition including any of the compounds ofthe present disclosure, where the administering is effective to providea therapeutically effective amount of the compound at the target site inthe subject.

The subject to be treated can be one that is in need of therapy, wherethe subject to be treated is one amenable to treatment using thecompounds disclosed herein. Accordingly, a variety of subjects may beamenable to treatment using the compounds disclosed herein. Generally,such subjects are “mammals”, with humans being of interest. Othersubjects can include domestic pets (e.g., dogs and cats), livestock(e.g., cows, pigs, goats, horses, and the like), rodents (e.g., mice,guinea pigs, and rats, e.g., as in animal models of disease), as well asnon-human primates (e.g., chimpanzees, and monkeys).

The present disclosure provides methods that include delivering acompound of the present disclosure to an individual having Alzheimer'sdisease, such as methods that include administering to the subject atherapeutically effective amount of a compound of the presentdisclosure. The methods are useful for treating a wide variety ofconditions and/or symptoms associated with Alzheimer's disease. In thecontext of Alzheimer's disease, the term “treating” includes one or more(e.g., each) of: reducing the severity of one or more symptoms,inhibiting the progression, reducing the duration of one or moresymptoms, and ameliorating one or more symptoms associated withAlzheimer's disease. In certain embodiments, methods of the presentdisclosure include administering a compound of the present disclosure toa subject, where the administering is effective for treating a diseasemediated through activity of the amylin receptor. In some instances,compounds of the present disclosure are effective for inhibiting theactivity of the amylin receptor.

The compounds described herein can be isolated by procedures known tothose skilled in the art. The compounds described herein may beobtained, for instance, by a resolution technique or by chromatographytechniques (e.g., silica gel chromatography, chiral chromatography,etc.). As used herein, the term “isolated” refers to compounds that arenon-naturally occurring and can be obtained or purified from syntheticreaction mixtures. Isolated compounds may find use in the pharmaceuticalcompositions and methods of treatment described herein.

The compounds described also include isotopically labeled compoundswhere one or more atoms have an atomic mass different from the atomicmass conventionally found in nature. Examples of isotopes that may beincorporated into the compounds disclosed herein include, but are notlimited to, ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, etc. Thus, thedisclosed compounds may be enriched in one or more of these isotopesrelative to the natural abundance of such isotope. By way of example,deuterium (²H; D) has a natural abundance of about 0.015%. Accordingly,for approximately every 6,500 hydrogen atoms occurring in nature, thereis one deuterium atom. Specifically contemplated herein are compoundsenriched in deuterium at one or more positions. Thus, deuteriumcontaining compounds of the disclosure have deuterium at one or morepositions (as the case may be) in an abundance of greater than 0.015%.In some embodiments, one or more (e.g., 1, 2, 3, 4, 5, 6, 7 or more)hydrogen atoms of a substituent group (e.g., an R-group) of any one ofthe subject compounds described herein are substituted with a deuterium.

Pharmaceutical Compositions

In certain embodiments, the disclosed compounds are useful for thetreatment of a disease or disorder, such as Alzheimer's disease.Accordingly, pharmaceutical compositions comprising at least onedisclosed compound are also described herein. For example, the presentdisclosure provides pharmaceutical compositions that include atherapeutically effective amount of a compound of the present disclosure(or a pharmaceutically acceptable salt or solvate or hydrate orstereoisomer thereof) and a pharmaceutically acceptable excipient.

A pharmaceutical composition that includes a subject compound may beadministered to a patient alone, or in combination with othersupplementary active agents. For example, one or more compoundsaccording to the present disclosure can be administered to a patientwith or without supplementary active agents. The pharmaceuticalcompositions may be manufactured using any of a variety of processes,including, but not limited to, conventional mixing, dissolving,granulating, dragee-making, levigating, emulsifying, encapsulating,entrapping, lyophilizing, and the like. The pharmaceutical compositioncan take any of a variety of forms including, but not limited to, asterile solution, suspension, emulsion, spray dried dispersion,lyophilisate, tablet, microtablets, pill, pellet, capsule, powder,syrup, elixir or any other dosage form suitable for administration.

A compound of the present disclosure may be administered to a subjectusing any convenient means capable of resulting in the desired reductionin disease condition or symptom. Thus, a compound can be incorporatedinto a variety of formulations for therapeutic administration. Moreparticularly, a compound can be formulated into pharmaceuticalcompositions by combination with appropriate pharmaceutically acceptableexcipients, carriers or diluents, and may be formulated intopreparations in solid, semi-solid, liquid or gaseous forms, such astablets, capsules, powders, granules, ointments, solutions,suppositories, injections, inhalants, aerosols, and the like.

Formulations for pharmaceutical compositions are described in, forexample, Remington's Pharmaceutical Sciences, by E. W. Martin, MackPublishing Co., Easton, Pa., 19th Edition, 1995, which describesexamples of formulations (and components thereof) suitable forpharmaceutical delivery of the disclosed compounds. Pharmaceuticalcompositions that include at least one of the compounds can beformulated for use in human or veterinary medicine. Particularformulations of a disclosed pharmaceutical composition may depend, forexample, on the mode of administration and/or on the location of thesubject to be treated. In some embodiments, formulations include apharmaceutically acceptable excipient in addition to at least one activeingredient, such as a compound of the present disclosure. In otherembodiments, other medicinal or pharmaceutical agents, for example, withsimilar, related or complementary effects on the disease or conditionbeing treated can also be included as active ingredients in apharmaceutical composition.

Pharmaceutically acceptable carriers useful for the disclosed methodsand compositions may depend on the particular mode of administrationbeing employed. In addition to biologically neutral carriers,pharmaceutical compositions to be administered can optionally containnon-toxic auxiliary substances (e.g., excipients), such as wetting oremulsifying agents, preservatives, and pH buffering agents, and thelike. The disclosed pharmaceutical compositions may be formulated as apharmaceutically acceptable salt of a disclosed compound.

In some embodiments, the disclosed pharmaceutical compositions may beformulated to cross the blood brain barrier (BBB). One strategy for drugdelivery through the blood brain barrier (BBB) entails disruption of theBBB, either by osmotic means such as mannitol or leukotrienes, orbiochemically by the use of vasoactive substances such as bradykinin. ABBB disrupting agent can be co-administered with the pharmaceuticalcompositions disclosed herein when the compositions are administered byintravenous injection. Other strategies to go through the BBB may entailthe use of endogenous transport systems, including carrier-mediatedtransporters such as glucose and amino acid carriers, receptor-mediatedtranscytosis for insulin or transferrin, and active efflux transporterssuch as p-glycoprotein. Active transport moieties may also be conjugatedto a compound disclosed herein for use in the methods disclosed hereinto facilitate transport across the epithelial wall of the blood vessel.Alternatively, drug delivery behind the BBB may be by intrathecaldelivery of therapeutics, e.g., administering the disclosedpharmaceutical compositions directly to the cranium, as through anOmmaya reservoir.

The term “unit dosage form,” as used herein, refers to physicallydiscrete units suitable as unitary dosages for human and animalsubjects, each unit containing a predetermined quantity of a compoundcalculated in an amount sufficient to produce the desired effect inassociation with a pharmaceutically acceptable diluent, excipient,carrier or vehicle. The specifications for a compound depend on theparticular compound employed and the effect to be achieved, and thepharmacodynamics associated with each compound in the subject.

The dosage form of a disclosed pharmaceutical composition may bedetermined by the mode of administration chosen. For example, inaddition to injectable fluids, topical or oral dosage forms may beemployed. Topical preparations may include eye drops, ointments, spraysand the like. Oral formulations may be liquid (e.g., syrups, solutionsor suspensions), or solid (e.g., powders, pills, tablets, or capsules).Methods of preparing such dosage forms are known, or will be apparent,to those skilled in the art.

Certain embodiments of the pharmaceutical compositions that include asubject compound may be formulated in unit dosage form suitable forindividual administration of precise dosages. The amount of activeingredient administered may depend on the subject being treated, theseverity of the affliction, and the manner of administration, and isknown to those skilled in the art. In certain instances, the formulationto be administered contains a quantity of the compounds disclosed hereinin an amount effective to achieve the desired effect in the subjectbeing treated.

Each therapeutic compound can independently be in any dosage form, suchas those described herein, and can also be administered in various ways,as described herein. For example, the compounds may be formulatedtogether, in a single dosage unit (that is, combined together in oneform such as capsule, tablet, powder, or liquid, etc.) as a combinationproduct. Alternatively, when not formulated together in a single dosageunit, an individual compound may be administered at the same time asanother therapeutic compound or sequentially, in any order thereof.

A disclosed compound can be administered alone, as the sole activepharmaceutical agent, or in combination with one or more additionalcompounds of the present disclosure or in conjunction with other agents.When administered as a combination, the therapeutic agents can beformulated as separate compositions that are administered simultaneouslyor at different times, or the therapeutic agents can be administeredtogether as a single composition combining two or more therapeuticagents. Thus, the pharmaceutical compositions disclosed hereincontaining a compound of the present disclosure optionally include othertherapeutic agents. Accordingly, certain embodiments are directed tosuch pharmaceutical compositions, where the composition further includesa therapeutically effective amount of an agent selected as is known tothose of skill in the art.

Methods of Administration

The subject compounds find use for treating a disease or disorder in asubject, such as Alzheimer's disease. The route of administration may beselected according to a variety of factors including, but not limitedto, the condition to be treated, the formulation and/or device used, thesubject to be treated, and the like. Routes of administration useful inthe disclosed methods include, but are not limited to, oral andparenteral routes, such as intravenous (iv), intraperitoneal (ip),rectal, topical, ophthalmic, nasal, intrathecal, and transdermal.Formulations for these dosage forms are described herein.

An effective amount of a subject compound may depend, at least, on theparticular method of use, the subject being treated, the severity of theaffliction, and the manner of administration of the therapeuticcomposition. A “therapeutically effective amount” of a composition is aquantity of a specified compound sufficient to achieve a desired effectin a subject (e.g., patient) being treated. For example, this may be theamount of a subject compound necessary to prevent, inhibit, reduce orrelieve a disease or disorder in a subject, such as Alzheimer's disease.Ideally, a therapeutically effective amount of a compound is an amountsufficient to prevent, inhibit, reduce or relieve a disease or disorderin a subject without causing a substantial cytotoxic effect on hostcells in the subject.

Therapeutically effective doses of a subject compound or pharmaceuticalcomposition can be determined by one of skill in the art. For example,in some instances, a therapeutically effective dose of a compound orpharmaceutical composition is administered with a goal of achievinglocal (e.g., tissue) concentrations that are at least as high as theEC₅₀ of an applicable compound disclosed herein.

The specific dose level and frequency of dosage for any particularsubject may be varied and may depend upon a variety of factors,including the activity of the subject compound, the metabolic stabilityand length of action of that compound, the age, body weight, generalhealth, sex and diet of the subject, mode and time of administration,rate of excretion, drug combination, and severity of the condition ofthe host undergoing therapy.

In some embodiments, multiple doses of a compound are administered. Thefrequency of administration of a compound can vary depending on any of avariety of factors, e.g., severity of the symptoms, condition of thesubject, etc. For example, in some embodiments, a compound isadministered once per month, twice per month, three times per month,every other week, once per week (qwk), twice per week, three times perweek, four times per week, five times per week, six times per week,every other day, daily (qd/od), twice a day (bds/bid), or three times aday (tds/tid), etc.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the present invention, and are not intended to limit thescope of what the inventors regard as their invention nor are theyintended to represent that the experiments below are all or the onlyexperiments performed. Efforts have been made to ensure accuracy withrespect to numbers used (e.g. amounts, temperature, etc.) but someexperimental errors and deviations should be accounted for. Unlessindicated otherwise, parts are parts by weight, molecular weight isweight average molecular weight, temperature is in degrees Celsius, andpressure is at or near atmospheric. By “average” is meant the arithmeticmean. Standard abbreviations may be used, e.g., bp, base pair(s); kb,kilobase(s); pl, picoliter(s); s or sec, second(s); min, minute(s); h orhr, hour(s); aa, amino acid(s); kb, kilobase(s); bp, base pair(s); nt,nucleotide(s); i.m., intramuscular(ly); i.p., intraperitoneal(ly); s.c.,subcutaneous(ly); and the like.

General Synthetic Procedures

Many general references providing commonly known chemical syntheticschemes and conditions useful for synthesizing the disclosed compoundsare available (see, e.g., Smith and March, March's Advanced OrganicChemistry: Reactions, Mechanisms, and Structure, Fifth Edition,Wiley-Interscience, 2001; or Vogel, A Textbook of Practical OrganicChemistry, Including Qualitative Organic Analysis, Fourth Edition, NewYork: Longman, 1978).

Compounds as described herein can be purified by any purificationprotocol known in the art, including chromatography, such as HPLC,preparative thin layer chromatography, flash column chromatography andion exchange chromatography. Any suitable stationary phase can be used,including normal and reversed phases as well as ionic resins. In certainembodiments, the disclosed compounds are purified via silica gel and/oralumina chromatography. See, e.g., Introduction to Modern LiquidChromatography, 2nd Edition, ed. L. R. Snyder and J. J. Kirkland, JohnWiley and Sons, 1979; and Thin Layer Chromatography, ed E. Stahl,Springer-Verlag, New York, 1969.

During any of the processes for preparation of the subject compounds, itmay be necessary and/or desirable to protect sensitive or reactivegroups on any of the molecules concerned. This may be achieved by meansof conventional protecting groups as described in standard works, suchas J. F. W. McOmie, “Protective Groups in Organic Chemistry”, PlenumPress, London and New York 1973, in T. W. Greene and P. G. M. Wuts,“Protective Groups in Organic Synthesis”, Third edition, Wiley, New York1999, in “The Peptides”; Volume 3 (editors: E. Gross and J. Meienhofer),Academic Press, London and New York 1981, in “Methoden der organischenChemie”, Houben-Weyl, 4^(th) edition, Vol. 15/1, Georg Thieme Verlag,Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, “Aminosauren, Peptide,Proteine”, Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982,and/or in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharide andDerivate”, Georg Thieme Verlag, Stuttgart 1974. The protecting groupsmay be removed at a convenient subsequent stage using methods known fromthe art.

The subject compounds, including compounds that are not commerciallyavailable, can be synthesized via a variety of different syntheticroutes using commercially available starting materials and/or startingmaterials prepared by conventional synthetic methods. A variety ofexamples of synthetic routes that can be used to synthesize thecompounds disclosed herein are described in the schemes below.

In certain embodiments, compounds of Formula (I) are synthesized usingmethods and conditions that are known to one of ordinary skill in theart, as depicted in Scheme 1:

wherein R, R¹, m, Q, W, X, Y, and Z are as defined herein.

The starting materials and reagents employed in Scheme 1 may be obtainedcommercially or through techniques known to one of ordinary skill in theart. Scheme 1 is an example of a method to generate compounds of Formula(I) where the exact steps and materials will depend on the functionalgroups present. The selection of the starting materials, reagent,substrates, base, protecting, solvent and leaving group can beaccomplished by one of ordinary skilled in the art. Regarding a nitrogenprotecting group, a non-limiting example is a t-butylcarbonate (Boc)group, which will allow protection of one nitrogen and let reactionoccur at the other nitrogen. Selective removal of the protecting groupis well documented in the literature and is of common knowledge to oneof ordinary skill in the art. For example, removal of a Boc group can beaccomplished with acid, such as trifluoroacetic acid or hydrochloricacid. For the leaving groups, some non-limiting examples are chloride,mesylate, and —O(C═O)OtBu, which will allow the desired nitrogen carbonbond to be formed. The selection of base will depend on the nature ofthe bond being created and functional groups present and somenon-limiting examples are: triethylamine, pyridine,N,N-diisopropylethylamine, sodium hydride, potassium tert-butoxide andpotassium carbonate.

In certain embodiments, compounds of Formula (II) are synthesized usingmethods and conditions that are known to one of ordinary skill in theart, as depicted in Scheme 2:

wherein R, R¹, m, Q, W, Y, and Z are as defined herein.

The starting materials and reagents employed in Scheme 2 may be obtainedcommercially or through techniques known to one of ordinary skill in theart. Scheme 2 is an example of a method to generate compounds of Formula(II) where the exact steps and materials will depend on the functionalgroups present. The selection of the starting materials, reagent,substrates, base, protecting, solvent and leaving group can beaccomplished by one of ordinary skilled in the art. Regarding a nitrogenprotecting group, a non-limiting example is a t-butylcarbonate (Boc)group, which will allow protection of one nitrogen and let reactionoccur at the other nitrogen. Selective removal of the protecting groupis well documented in the literature and is of common knowledge to oneof ordinary skill in the art. For example, removal of a Boc group can beaccomplished with acid, such as trifluoroacetic acid or hydrochloricacid. For the leaving groups, some non-limiting examples are chloride,mesylate, and —O(C═O)OtBu, which will allow the desired nitrogen carbonbond to be formed. The selection of base will depend on the nature ofthe bond being created and functional groups present and somenon-limiting examples are: triethylamine, pyridine,N,N-diisopropylethylamine, sodium hydride, potassium tert-butoxide andpotassium carbonate.

In certain embodiments, compounds of Formula (III) are synthesized usingmethods and conditions that are known to one of ordinary skill in theart, as depicted in Scheme 3:

wherein R, R¹, m, W and Z are as defined herein.

The starting materials and reagents employed in Scheme 3 may be obtainedcommercially or through techniques known to one of ordinary skill in theart. Scheme 3 is an example of a method to generate compounds of Formula(III) where the exact steps and materials will depend on the functionalgroups present. The selection of the starting materials, reagent,substrates, base, protecting, solvent and leaving group can beaccomplished by one of ordinary skilled in the art. Regarding a nitrogenprotecting group, a non-limiting example is a t-butylcarbonate (Boc)group, which will allow protection of one nitrogen and let reactionoccur at the other nitrogen. Selective removal of the protecting groupis well documented in the literature and is of common knowledge to oneof ordinary skill in the art. For example, removal of a Boc group can beaccomplished with acid, such as trifluoroacetic acid or hydrochloricacid. For the leaving groups, some non-limiting examples are chloride,mesylate, and —O(C═O)OtBu, which will allow the desired nitrogen carbonbond to be formed. The selection of base will depend on the nature ofthe bond being created and functional groups present and somenon-limiting examples are: triethylamine, pyridine,N,N-diisopropylethylamine, sodium hydride, potassium tert-butoxide andpotassium carbonate.

In certain embodiments, compounds of Formula (IV) are synthesized usingmethods and conditions that are known to one of ordinary skill in theart, as depicted in Scheme 4:

wherein R, R¹, m, W and Z are as defined herein.

The starting materials and reagents employed in Scheme 4 may be obtainedcommercially or through techniques known to one of ordinary skill in theart. Scheme 4 is an example of a method to generate compounds of Formula(IV) where the exact steps and materials will depend on the functionalgroups present. The selection of the starting materials, reagent,substrates, base, protecting, solvent and leaving group can beaccomplished by one of ordinary skilled in the art. Regarding a nitrogenprotecting group, a non-limiting example is a t-butylcarbonate (Boc)group, which will allow protection of one nitrogen and let reactionoccur at the other nitrogen. Selective removal of the protecting groupis well documented in the literature and is of common knowledge to oneof ordinary skill in the art. For example, removal of a Boc group can beaccomplished with acid, such as trifluoroacetic acid or hydrochloricacid. For the leaving groups, some non-limiting examples are chloride,mesylate, and —O(C═O)OtBu, which will allow the desired nitrogen carbonbond to be formed. The selection of base will depend on the nature ofthe bond being created and functional groups present and somenon-limiting examples are: triethylamine, pyridine,N,N-diisopropylethylamine, sodium hydride, potassium tert-butoxide andpotassium carbonate.

In certain embodiments, compounds of Formula (V), (VI), and (VII) aresynthesized using methods and conditions that are known to one ofordinary skill in the art, as depicted in Scheme 5:

wherein R⁷ is defined herein.

The starting materials and reagents employed in Scheme 5 may be obtainedcommercially or through techniques known to one of ordinary skill in theart and documented in the examples herein. Scheme 5 is an example ofmethods to generate compounds of Formula (V), Formula (VI) and Formula(VII) where the exact steps and materials will depend on the functionalgroups present. The selection of the starting materials, reagent,substrates, base, protecting, solvent and leaving group can beaccomplished by one of ordinary skilled in the art. For an oxygenprotecting group, a non-limiting example is a t-butyldimethylsilylgroup, which will allow protection of the alcohol oxygen. Selectiveremoval of the protecting group is well documented in the literature andis of common knowledge to one of ordinary skill in the art. For example,removal of a t-butyldimethylsilyl group can be accomplished withfluoride, such as tetrabutylammonium fluoride or pyridinium hydrogenfluoride or KF. For the leaving groups, some non-limiting examples arechloride, mesylate, and —O(C═O)OtBu, which will allow the desirednitrogen carbon bond to be formed. The selection of base will depend onthe nature of the bond being created and functional groups present andsome non-limiting examples are: triethylamine, pyridine,N,N-diisopropylethylamine, sodium hydride, potassium tert-butoxide andpotassium carbonate.

Schemes 1, 2, 3, 4 and 5 are meant to be by way of non-limiting examplesonly, and one of ordinary skill in the art will understand thatalternate reagents, solvents or starting materials can be used to makecompounds of Formula (I) and/or (II) and/or (III) and/or (IV) and/or (V)and/or (VI) and/or (VII) and/or other compounds contained herein.

Example 1: Synthesis of Compounds

All reagents and solvents were used as purchased from commercialsources. Moisture sensitive reactions were carried out under a nitrogenatmosphere. Reactions were monitored by TLC using pre-coated silica gelaluminum plates containing a fluorescent indicator (F-254). Detectionwas done with UV (254 nm). Alternatively, the progress of a reaction wasmonitored by LC/MS. Specifically, but without limitation, the followingabbreviations were used, in addition to the other ones described herein,in the examples: Bn (benzyl); Boc (tert-butoxycarbonyl); Boc₂O(di-tert-butyl dicarbonate); cat. (catalytic amount); DCM(dichloromethane); dioxane (1,4-dioxane); DIPEA(N,N-diisopropylethylamine); DMAP (4-dimethylaminopyridine); DMF(N,N-dimethylformamide); EDCI (N-ethyl-N′-carbodiimide); EtOH (ethanol);ether or Et₂O (diethyl ether); Et₃N (triethylamine); HATU(1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxide hexafluorophosphate orN-[(dimethylamino)-1H-1,2,3-triazolo-[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminiumhexafluorophosphate N-oxide); hex (hexanes); MeCN (acetonitrile); MeOH(methanol); μW (microwave); MS (molecular sieves); O/N (overnight);Pd(PPh₃)₄ (tetrakis(triphenylphosphine) palladium(0)); Pd/C (palladiumon carbon); Pd(OH)₂/C (palladium hydroxide on carbon); RT or rt (room orambient temperature); TBS (tert-butyldimethylsilyl); TfOH(trifloromethane sulfonic acid); THF (tetrahydrofuran); TMS(trimethylsilyl). ¹H NMR spectra were recorded at RT with a BrukerAvanche III 600 MHz NMR spectrometer equipped with a Bruker's 5 mm PABBOprobe. Chemical shifts are reported in ppm downfield fromtetramethylsilane using residual solvent signals as internal reference.NMR data were processed utilizing ACD/Spectrus processor (v2016.1.1,ACD/Labs Inc.). Nomenclature for the naming of compounds, such as forCompound Examples and intermediate compounds, were performed usingACD/Name (Chemists' Version from ACD/Labs Inc.) to generate theIUPAC-style names. Naming of commercial or literature compounds utilizedSciFinder, ACD/Names, and common or trivial names known to those skilledin the art.

Microwave assisted reactions were performed using an Anton Paar“Monowave 200” Microwave Synthesis Reactor with magnetron power 850 W.Unless stated otherwise the temperature was reached as fast as possibleand controlled by built-in IR sensor (temperature uncertainty ±5° C.).Reaction was carried out either in 10 mL or 30 mL vials, with thedefault stirrer speed 600 rpm.

The LC/MS system used for monitoring the progress of reactions,assessing the purity (absorbance at 254 nm) and identity of the productconsisted of Dionex ULTIMATE 3000 uHPLC module and Thermo Scientific LTQXL mass-spectrometer with electrospray ionization and Ion-Trap type ofdetector (alternating positive-negative mode). Separation was performedwith Thermo Scientific™ Accucore™ aQ C18 Polar Endcapped LC column (100mm×2.1 mm; particle size 2.6 m, 80 Å). The column was maintained at 30°C. Commercial HPLC-grade methanol, acetonitrile and domestic ‘millipore(Milli-Q)’ water used for chromatography were modified by adding 0.1%(v/v) of formic acid. The eluent was delivered with constant flow rateof 0.4 mL/min, column was equilibrated for 5 min with the correspondingeluent prior to injection of the sample (1 μL) and one of the followingseparation conditions were used:

Eluent Systems:

A—Gradient of MeOH-Water, 15% to 65% in 5 min, 65% to 95% in 2.5 min,followed by 4 min of isocratic MeOH-water 95%;

B—Gradient of Methanol-Water, 30 to 65% in 4.75 min, then to 95% in 2.5min, followed by 4 min of isocratic MeOH-water 95%;

C—Gradient of MeOH-Water, 10% to 45% in 5 min, 45% to 95% in 2.5 min,followed by 4 min of isocratic MeOH-water 95%; and

D—Gradient of Methanol-Water, 45 to 95% in 5.25 min, followed by 5 minof isocratic MeOH-water 95%.

Compound 1 Synthesis of1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 1

Compound 1 was synthesized as in Scheme 6.

Preparation of 3-(2-nitroanilino)propan-1-ol, (2). To a solution of1-fluoro-2-nitrobenzene (1) (60.0 g, 430 mmol) in DMF (200 mL) was addedK₂CO₃ (178.0 g, 1.30 mol), and after cooling in an ice bath3-amino-1-propanol (48.0 g, 640 mmol) via a dropping funnel. Afterovernight, the reaction mixture was vacuum-filtered and the solid waswashed with EtOAc (2×100 mL). The solvent was removed under reducedpressure. The crude residue was dissolved in EtOAc (500 mL). The organicsolution was washed thoroughly with water (3×150 mL) and brine (200 mL).The organic phase was dried over Na₂SO₄, filtered and concentrated underreduced pressure to dryness. To the residue was added hexanes (300 mL)and after 30 mins, to break the solid into fine powder, the product wascollected by vacuum filtration, which provided (2) (81.4 g, 97% yield)as an orange solid.

Preparation of 3-(2-aminoanilino)propan-1-ol, (3). To a 1 Lround-bottomed flask was added (2) (20.0 g, 102.0 mmol) and MeOH (300mL). The mixture was degassed and put under a nitrogen atmosphere.Carefully 10% Pd/C (2.2 g) was added and after degassing 3 times withnitrogen, a balloon filled with hydrogen gas was added. The balloon wasrefilled with hydrogen gas as needed. After stirring for 6 h, thereaction mixture was filtered through a pad of Celite®, which was washedwith MeOH (200 mL). The filtrate was concentrated under reduced pressureproviding (3) (16.6 g, 98% yield) as a brown sticky oil. A second batch(50 g, 254.8 mmol) was done with the same procedure, which provided (3)(41.7 g, 99% yield). The material was used in the next step with outfurther purification.

Preparation of 1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 1.To a solution of 3-(2-aminoanilino)propan-1-ol (3) (16.6 g, 100 mmol)and Et₃N (71.0 mL, 510 mmol) in DCM (400 mL) cooled in an ice bath wasslowly added a solution of ethyl chlorooxoacetate (16.8 mL, 150 mmol) inDCM (50 mL) via a dropping funnel. After overnight, distilled water (100mL) was added and after stirring for 30 min the mixture was filtered. Asecond batch with 41.7 g (251 mmol) of 3-(2-aminoanilino)propan-1-ol (3)was done using the same procedure. The two batches were combined forpurification. The solids from both batches were washed with water andair dried after which they were combined and dissolved in a mixture ofDCM and MeOH (1:1, 4 L). To the solution was added activated charcoal(50 g). After filtration through a pad of Celite®, the filtrate wasconcentrated to a volume of about 300 mL when solid precipitated fromthe solution. The suspension was filtered and collected solid driedproviding a brown solid (13 g). The mother liquor from the firstfiltration was concentrated to a volume of 400 mL placed in the fridgeovernight. The resulting solid was collected by gravimetric filtrationand after drying provided a brown solid (20 g). The solids (13 g and 20g) were dissolved in DCM and MeOH. It required approximately 1 L of DCMand 1 L of MeOH to fully dissolve 13 g of product. To the resultingsolution was added activated charcoal and the mixture was warmed toreflux for about 30 min. Then it was filtered carefully through a pad ofCelite® while the solution was still warm. The filtrate was thenconcentrated until precipitation appeared. The solid (33 g) was combinedfrom the separate recrystallizations collected and the filtrates fromall previous steps were also combined, concentrated under reducedpressure and dried before being re-dissolved in DCM (1.2 L) and MeOH(1.2 L). To the resulting solution was added K₂CO₃ (76.2 g, 551 mmol)and after overnight the salts were filtered through a pad of Celite® andthe filtrate was absorbed to silica gel (200 g). The silica gel wasdried completely under the vacuum. The silica gel with the crude productwas loaded on a silica gel column and the product eluted with a gradientof MeOH/CHCl₃, 5 to 20%. The pure fractions were combined and treatedwith activated charcoal, filtered through a pad of Celite® andconcentrated to a volume when solid appeared. The suspension wasfiltered providing an off white solid (11 g). The purified solids (33 gand 11 g) were combined and Milli-Q water (300 mL) was added. Thesuspension was heated to reflux for 2 h. The water was removed underreduced pressure and dried completely, which provided 1 (43.7 g, 57%yield) as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.03 (br s, 1H), 7.48-7.33 (m, 1H),7.26-7.04 (m, 3H), 4.66 (t, J=5.1 Hz, 1H), 4.27-4.07 (m, 2H), 3.52 (q,J=5.6 Hz, 2H), 1.83-1.72 (m, 2H). LC/MS: Eluent system C (retentiontime: 4.35 min); ESI-MS 221 [M+H]⁺.

Compound 2 Synthesis of1-(3-methoxypropyl)-4-methyl-1,4-dihydroquinoxaline-2,3-dione, 2

Compound 2 was synthesized as in Scheme 7.

Preparation of1-(3-methoxypropyl)-4-methyl-1,4-dihydroquinoxaline-2,3-dione, 2. To asolution of 1-methyl-1,4-dihydroquinoxaline-2,3-dione (4) (100 mg, 0.57mmol) in DMF (5 mL) was added NaH (114 mg, 2.85 mmol, 60% in oil). After15 min, to the mixture was added 1-bromo-3-methoxypropane (5) (0.19 mL,2.85 mmol). The mixture was warmed in an oil bath at 100° C. Afterovernight, the volatiles were removed under reduced pressure. Theresulting mixture was dissolved in DCM (10 mL), silica gel (5 g) wasadded and the solvent was removed under reduced pressure. The resultingdried silica gel with mixture absorbed was loaded on a silica gel columnand the compound was eluted with a gradient of MeOH/CHCl₃, 0 to 5%,which generated 2 (22 mg, 16% yield) as a pale pink solid.

¹H NMR (600 MHz, DMSO-d₆) δ 7.53-7.41 (m, 2H), 7.34-7.23 (m, 2H),4.21-4.17 (m, 2H), 3.55 (s, 3H), 3.43 (t, J=6.2 Hz, 2H), 3.25 (s, 3H),1.94-1.79 (m, 2H). LC/MS: Eluent system A (retention time: 5.66 min);ESI-MS 249 [M+H]⁺.

Compound 3 Synthesis of1-(3-hydroxypropyl)-4-methyl-1,4-dihydroquinoxaline-2,3-dione, 3

Compound 3 was synthesized as in Scheme 8.

Preparation of1-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4-methyl-1,4-dihydroquinoxaline-2,3-dione(7). To a solution of 1-methyl-1,4-dihydroquinoxaline-2,3-dione (4) (100mg, 0.57 mmol) in DMF (5 mL) was added NaH (114 mg, 2.85 mmol, 60% inoil). After stirring for 15 min, to the mixture was added3-(tert-butyldimethylsilyloxy)propyl bromide (6) (0.26 mL, 1.14 mmol).The mixture was heated in an oil bath at 100° C. After overnight, thesolvent was removed under reduced pressure. The resulting material wasdissolved in DCM (10 mL), silica gel (5 g) was added and the solvent wasremoved under reduced pressure. The resulting dried silica gel withabsorbed mixture was loaded on a silica gel column and the compound waseluted with a gradient of MeOH/CHCl₃, 0 to 5%, which generated (7) (128mg, 64% yield) as a white solid.

Preparation of1-(3-hydroxypropyl)-4-methyl-1,4-dihydroquinoxaline-2,3-dione, 3. To asolution of1-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4-methyl-1,4-dihydroquinoxaline-2,3-dione(7) (128 mg, 0.37 mmol) in CH₃CN (5 mL) was added KF (22 mg 0.37 mmol),followed by TMSCl (0.047 mL, 0.37 mmol) and 1 drop of water. After 1 h,the reaction mixture was quenched with a saturated NaHCO₃ aqueoussolution. The mixture was extracted with DCM (3×15 mL). The combinedorganic phase was washed with brine (15 mL), dried over Na₂SO₄, filteredand concentrated under reduced pressure. The product was purified bycolumn chromatography on silica gel with a gradient of MeOH/EtOAc, 0 to10%, which generated 3 as a white solid (61 mg, 70% yield).

¹H NMR (600 MHz, DMSO-d₆) δ 7.54-7.47 (m, 1H), 7.46-7.41 (m, 1H),7.38-7.19 (m, 2H), 4.68 (t, J=5.1 Hz, 1H), 4.23-4.17 (m, 2H), 3.55 (s,3H), 3.54-3.51 (m, 2H), 1.83-1.75 (m, 2H). LC/MS: Eluent system A(retention time: 4.05 min); ESI-MS 235 [M+H]⁺.

Compound 4 Synthesis of1-(3-methoxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 4

Compound 4 was synthesized as in Scheme 9.

Preparation of 1-(3-methoxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 4.To a solution of 1,4-dihydroquinoxaline-2,3-dione (8) (1.0 g, 6.2 mmol)in DMF (5 mL) was added NaH (74 mg, 1.86 mmol, 60% in oil). After 30min, to the mixture was added 1-bromo-3-methoxypropane (5) (0.07 mL,0.62 mmol). The resulting mixture was warmed to 100° C. in an oil bath.After overnight, the oil bath was removed and after cooling to ambienttemperature methanol was slowly added. The volatiles were removed underreduced pressure. The resulting material was dissolved in DCM/MeOH(1:1,20 mL), silica gel (10 g) was added and the solvent was removed underreduced pressure. The resulting dried silica gel with material absorbedwas loaded on a silica gel column and the compound was eluted with agradient of MeOH/CHCl₃, 0 to 20%, which generated 4 (34 mg, 23% yield)as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.03 (br s, 1H), 7.44-7.34 (m, 1H),7.26-7.09 (m, 3H), 4.21-4.10 (m, 2H), 3.42 (t, J=6.2 Hz, 2H), 3.25 (s,3H), 1.92-1.81 (m, 2H). LC/MS: Eluent system A (retention time: 6.30min); ESI-MS 235 [M+H]⁺.

Compound 5 Synthesis of1-(3-phenoxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 5

Compound 5 was synthesized as in Scheme 10.

Preparation of 1-(3-phenoxypropyl)-1,4-dihydroquinoxaline-2,3-dione 5.To a solution of 1,4-dihydroquinoxaline-2,3-dione (8) (1.0 g, 6.2 mmol)in DMF (5 mL) was added NaH (74 mg, 1.86 mmol, 60% in oil). After 30min, to the mixture was added 3-phenoxypropyl bromide (9) (0.10 mL, 0.62mmol). The resulting mixture was warmed to 100° C. in an oil bath. Afterovernight, the oil bath was removed and after cooling to ambienttemperature methanol was slowly added. The volatiles were removed underreduced pressure. The resulting material was dissolved in DCM/MeOH (1:1,20 mL), silica gel (5 g) was added and the solvent was removed underreduced pressure. The resulting dried silica gel with material absorbedwas loaded on a silica gel column and the compound was eluted with agradient of MeOH/CHCl₃, 0 to 10%, which generated 5 (103 mg, 56% yield)as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.04 (brs, 1H), 7.46-7.41 (m, 1H),7.32-7.27 (m, 2H), 7.23-7.14 (m, 3H), 6.97-6.91 (m, 3H), 4.32-4.25 (m,2H), 4.09 (t, J=6.2 Hz, 2H), 2.12-2.05 (m, 2H). LC/MS: Eluent system A(retention time: 7.85 min); ESI-MS 297 [M+H]⁺.

Compound 6 Synthesis of3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl acetate, 6

Compound 6 was synthesized as in Scheme 11.

Preparation of 4-(3-bromopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one,(11). A solution of methyl bromoacetate (15 g, 0.098 mol) in THF (30 mL)was cooled in a −78° C. bath and to this was added a precooled to thesame temperature solution of o-phenylenediamine (10) (10.0 g, 0.0925mol) in THF (50 mL), containing potassium carbonate (15.0 g, 0.108 mol).After thoroughly stirring the mixture, it was left at −20° C. in thefreezer. After overnight, the mixture was warmed to ambient temperature.After 1 h, the reaction flask was placed in the ice-salt bath and onceinternal temperature reached −5° C. the ice-cold solution of3-bromopropanoyl chloride (17.1 g, 0.099 mol) in THF (15 mL) was addedslowly so that the temperature of the reaction mixture didn't raiseabove 10° C. After 30 minutes, the ice-salt bath was removed and afterone hour at room temperature the mixture was partitioned between ethylacetate (200 mL) and water (100 mL). The organic layer was dried oversodium sulfate, filtered, and concentrated under reduced pressure. Theproduct was purified by column chromatography on silica gel (eluted withhexane-DCM-ethyl acetate 2:1:0-1:1:2), which provided (11) (9.4 g, 36%over two steps) as a brownish powder.

Preparation of 3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propylacetate, 6. A solution of4-(3-bromopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one (11) (660 mg, 2.33mmol), sodium acetate (300 mg, 3.66 mmol) in glacial acetic acid (8 mL)in a 30 mL microwave vial was heated under microwave irradiation for 4hours with the temperature set at 125° C. Another portion of sodiumacetate (300 mg, 3.66 mmol) was then added and the mixture was heatedfor additional 4 hours at 125° C. The mixture was concentrated underreduced pressure, partitioned between water and chloroform and theorganic layer was separated and concentrated. A DCM solution of theresulting residue was loaded on silica gel. Purification wasaccomplished by silica gel column chromatography (eluted withhexanes-ethyl acetate 35%→70%) provided two products:4-acryloyl-3,4-dihydroquinoxalin-2(1H)-one (12) (85 mg, 18% yield) as aby-product and the target 6 (150 mg, 25% yield) as a white powder.

¹H NMR (600 MHz, DMSO-d₆) δ 10.71 (s, 1H), 7.49 (br s, 1H), 7.22 (br s,1H), 7.08-7.01 (m, 2H), 4.35 (s, 2H), 4.26-4.18 (m, 2H), 2.88 (br s,2H), 1.95 (s, 3H). LC/MS: Eluent system C (retention time: 6.17 min);ESI-MS 263 [M+H]⁺ and 261 [M−H]⁻.

Compound 7 Synthesis of4-(3-((2-hydroxyethyl)amino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one,7

Compound 7 was synthesized as in Scheme 12.

Preparation of4-(3-((2-hydroxyethyl)amino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one,7. To a solution of 4-acryloyl-3,4-dihydroquinoxalin-2(1H)-one (12) (40mg, 0.2 mmol) in methanol (5 mL) was added ethanolamine (0.50 g, 8.2mmol). After 30 min, the mixture was concentrated under pressure and theproduct purified by silica gel column chromatography (eluted with ethylacetate-methanol 2%→50%), which after trituration with ether provided 7(16 mg, 30% yield) as a white powder.

¹H NMR (600 MHz, DMSO-d6) δ 10.68 (br s, 1H), 7.50 (br s, 1H), 7.23-7.17(m, 1H), 7.08-7.00 (m, 2H), 4.42 (s, 1H), 4.34 (s, 2H), 2.77-2.71 (m,2H), 2.67-2.62 (m, 2H), 2.52-2.44 (m, 4H). LC/MS: Eluent system C(retention time: 1.16 min); ESI-MS 264 [M+H]⁺ and 262 [M−H]⁻.

Compound 8 Synthesis of4-(3-(dimethylamino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one, 8

Compound 8 was synthesized as in Scheme 13.

Preparation of4-(3-(dimethylamino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one, 8. To asolution of 4-acryloyl-3,4-dihydroquinoxalin-2(1H)-one (12) (40 mg, 0.2mmol) in methanol (5 mL) was added a 2N solution of dimethylamine in THF(2 mL, 4 mmol). After 30 min, the mixture was concentrated under reducedpressure and the product was purified by silica gel columnchromatography (eluted with ethyl acetate-methanol 2%→50%), whichproduced 8 (30 mg, 61% yield) as a yellow-brown glass like stickyliquid.

¹H NMR (600 MHz, DMSO-d6) δ 10.60 (br s, 1H), 7.65-7.36 (m, 1H),7.28-6.97 (m, 3H), 4.34 (s, 2H), 2.77-2.40 (m, 4H), 2.03 (br s, 6H).LC/MS: Eluent system C (retention time: 1.09 min); ESI-MS 248 [M+H]⁺.

Compound 9 Synthesis of1-(2-hydroxyethyl)-1,4-dihydroquinoxaline-2,3-dione, 9

Compound 9 was synthesized as in Scheme 14.

Preparation of1-(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)-1,4-dihydroquinoxaline-2,3-dione(13). To a solution of 1,4-dihydroquinoxaline-2,3-dione (8) (1.0 g, 6.2mmol) in DMF (10 mL) was added NaH (248 mg, 6.2 mmol, 60% in oil). After30 min, to the mixture was added 2-(tert-butyldimethylsilyloxy)ethylbromide (0.13 mL, 0.62 mmol). The resulting mixture was heated in an oilbath at 100° C. After overnight, the solvent was removed under reducedpressure. The crude material was dissolved in DCM/MeOH (1:1, 20 mL),silica gel (10 g) was added and the volatiles were removed under reducedpressure. The resulting dried silica gel with the mixture absorbed wasloaded on silica gel column and the product was eluted with a gradientof MeOH/CHCl₃, 0 to 10%, which generated (13) (122 mg, 61% yield) as awhite solid.

Preparation of 1-(2-hydroxyethyl)-1,4-dihydroquinoxaline-2,3-dione, 9.To a solution of1-(2-{[tert-butyl(dimethyl)silyl]oxy}ethyl)-1,4-dihydroquinoxaline-2,3-dione(13) (122 mg, 0.38 mmol) in CH₃CN (10 mL) was added KF (22 mg, 0.38mmol), followed by TMSCl (0.048 mL, 0.38 mmol) and 1 drop of water.After 1 h, the reaction mixture was quenched with a saturated NaHCO₃aqueous solution. The resulting mixture was extracted with DCM (3×15mL). The combined organic phase was washed with brine (15 mL), driedover Na₂SO₄, filtered and concentrated under reduced pressure. Theproduct was purified by column chromatography on silica gel with agradient of MeOH/CHCl₃, 0 to 20%, which generated 9 as a white solid (65mg, 83% yield).

¹H NMR (600 MHz, DMSO-d₆) δ 12.14 (br s, 1H), 7.62-7.52 (m, 1H),7.36-7.21 (m, 3H), 4.99 (t, J=6.0 Hz, 1H), 4.32 (t, J=6.2 Hz, 2H), 3.78(q, J=6.0 Hz, 2H). LC/MS: Eluent system C (retention time: 3.11 min);ESI-MS 207 [M+H]⁺.

Compound 10 Synthesis of4-(3-hydroxypropanoyl)-3,4-dihydroquinoxalin-2(1H)-one, 10

Compound 10 was synthesized as in Scheme 15

Preparation of 3-(benzyloxy)propanoyl chloride, (15). An oven-dried, 1.0L round-bottomed flask equipped with rubber septum containingthermometer, nitrogen inlet and outlet, cooling bath, and magneticstirrer, was charged with 3-(benzyloxy)propanoic acid (14) (27.5 g,0.153 mol) and anhydrous DCM (500 mL). The solution was cooled to 0-5°C. (internal temperature) using an ice-water bath. Oxalyl chloride (25.8mL, 0.305 mol) was then added dropwise over 10 min under nitrogenatmosphere. After 15 min at 0-5° C., anhydrous DMF (0.75 mL) was addeddropwise over 15 min. After 15 min, the ice bath was removed and thereaction warmed to room temperature. After 1 h at ambient temperature,the mixture was concentrated under reduced pressure using a 30° C. waterbath, it was then co-evaporated with anhydrous DCM (3×100 mL), and driedunder reduced pressure using a 35° C. water bath for 1.0 h, whichgenerated (15) (30.32 g, quantitative yield) as a pinkish-orange colorsolid. This was used in the next step without further purification.

Preparation of4-[3-(benzyloxy)propanoyl]-3,4-dihydroquinoxalin-2(1H)-one, (17). To3-(benzyloxy)propanoyl chloride (15) (30.32 g, 0.153 mol) in a 1.0 Lround-bottomed flask, was added solid 3,4-dihydroquinoxalin-2(1H)-one(16) (20.35 g, 0.137 mol) and anhydrous DMF (300 mL). After 15 min,anhydrous NaHCO₃ (15.4 g, 0.183 mol) was added in one portion. Afterovernight, a saturated aqueous NaHCO₃ solution (100 mL) was added andsaturated brine solution (500 mL), the resulting mixture was extractedwith CHCl₃ (3×750 mL). The combined organics were dried over anhydrousMg₂SO₄, filtered, and concentrated under reduced pressure using a 50° C.water bath. The resulting residue was dissolved in CHCl₃ (200 mL) andadsorbed on silica gel (50 g). The product was then purified by Biotage®(330 g Silicycle column) eluting with a gradient of 0% to 6%,MeOH:CHCl₃. The resulting tan color solid was dissolved in CHCl₃:MeOH(90:10, 500 mL) and activated charcoal (30 g) was added. After 1 h, thecharcoal was removed by vacuum filtration through a bed of Celite®, andthe bed was washed with CHCl₃:MeOH (90:10, 3×200 mL). The combinedfiltrates were concentrated under reduced pressure that afforded (17) asan off white solid (37.6 g, 79% yield).

Preparation of 4-(3-hydroxypropanoyl)-3,4-dihydroquinoxalin-2(1H)-one,10. To a solution of4-[3-(benzyloxy)propanoyl]-3,4-dihydroquinoxalin-2(1H)-one (17) (32.6 g,0.105 moles) in THF (300.0 mL) was added 10% Pd(OH)₂/C (3.26 g). Twoballoons were filled with hydrogen gas and affixed onto theround-bottomed flask. After 5 h, the resulting mixture was filteredusing vacuum filtration through a bed of Celite®, and the bed was washedthoroughly with CHCl₃:MeOH (80:20, 5×250 mL). The combined filtrateswere concentrated under reduced pressure. Similarly, another batch ofhydrogenation was carried with 5.0 g (16.11 mmol) of (17) and 0.5 g of10% Pd(OH)₂/C in 50 mL THF. The crude from the two batches were combinedand adsorbed on silica gel (50 g) using CHCl₃:MeOH (80:20, 200 mL). Theproduct was purified by Biotage® (330 g silicycle HP column) elutingwith a gradient of 0% to 20%, MeOH:CHCl₃. To the resulting solid wasadded 300 mL water and the resulting mixture was refluxed for 1 h andthen after cooling to ambient temperature was concentrated under reducedpressure with a water bath at 50° C., which afforded 10 as an off-whitesolid (23.6 g, 88% yield).

¹H NMR (600 MHz, DMSO-d₆) δ 10.69 (s, 1H), 7.53 (br s, 1H), 7.24-7.18(m, 1H), 7.06-7.01 (m, 2H), 4.64 (t, J=5.2 Hz, 1H), 4.35 (s, 2H), 3.67(q, J=6.0 Hz, 2H), 2.66 (br. t, J=5.8 Hz, 2H). LC/MS: Eluent system C(retention time: 4.17 min); ESI-MS: 221 [M+H]⁺.

Compounds 11 and 12 Synthesis of1-(3-hydroxypropyl)-4-methoxy-1,3-dihydro-2H-1,5-benzodiazepin-2-one, 11and 1-(3-hydroxypropyl)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione, 12

Compounds 11 and 12 were synthesized as in Scheme 16.

Preparation of1-(3-hydroxypropyl)-4-methoxy-1,3-dihydro-2H-1,5-benzodiazepin-2-one, 11and 1-(3-hydroxypropyl)-1H-1,5-benzodiazepine-2,4(3H,5H)-dione, 12. Amixture of 3-(2-aminoanilino)propan-1-ol (2), (100 mg, 0.60 mmol) anddiethyl malonate (18) (1.0 mL, 6.62 mmol) were put in a dry 10 mLmicrowave vial and the vial was positioned in a microwave reactor thatwas then set 200° C. After 30 min, the reaction mixture was cooled toambient temperature and methanol (3 mL) and K₂CO₃ (300 mg, 2.17 mmol)were added. After overnight, the mixture was vacuum filtered and thesolid was washed with CHCl₃:MeOH (9:1, 3×10 mL). The combined filtrateswere concentrated under reduced pressure. The mixture was adsorbed onsilica gel (5 g) using CHCl₃ (5 mL) and purified by column purificationon silica gel (Biotage®, 12 g silicycle column, eluted withchloroform-methanol: 0% to 2%), which afforded 11 as an off-white solid(33.9 mg, 23% yield).

¹H NMR (600 MHz, DMSO-d₆) δ 7.57 (d, J=7.9 Hz, 1H), 7.55 (d, J=7.9 Hz,1H) 7.24-7.22 (m, 1H), 7.19-7.17 (m, 1H), 4.71 (t, J=4.9 Hz, 1H), 4.26(t, J=7.2 Hz, 2H), 4.13 (s, 2H), 3.67 (s, 3H), 3.40-3.38 (m, 2H),1.92-1.81 (m, 2H). LC/MS: Eluent system C (retention time: 2.75 min);ESI-MS: 249 [M+H]⁺.

Further elution of the column with a gradient of 2% to 10% methanol inchloroform produced 12, which was triturated with EtOAc and hexanes toafford pure 12 as a white solid (17.8 mg, 13% yield).

¹H NMR (600 MHz, DMSO-d₆) δ 10.39 (s, 1H), 7.58-7.52 (m, 1H), 7.30-7.23(m, 2H), 7.21-7.15 (m, 1H), 4.44 (t, J=5.1 Hz, 1H), 4.27-4.17 (m, 1H),3.76-3.64 (m, 1H), 3.40-3.36 (m, 1H), 3.31-3.21 (m, 2H), 3.01-2.88 (m,1H), 1.64-1.43 (m, 2H). LC/MS: Eluent system C (retention time: 4.40min); ESI-MS: 235 [M+H]⁺.

Compound 13 Synthesis of1-(3-hydroxypropyl)-6-(pyridin-3-yl)-1,4-dihydroquinoxaline-2,3-dione,13

Compound 13 was synthesized as in Scheme 17.

Preparation of 2-nitro-4-(pyridin-3-yl)aniline, (21). A mixture of4-bromo-2-nitroaniline (19) (400 mg, 1.84 mmol),4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborolane (20) (450 mg, 2.19mmol), water (1 mL), K₂CO₃ (304.1 mg, 2.2 mmol) and DMF (10 mL) waspurged with N₂ for 5 min. To this mixture was added Pd(PPh₃)₄ (106.3 mg,0.092 mmol) and the resulting mixture was heated in an oil bath at 120°C. After 1 h, the mixture was concentrated under reduced pressure andthe residue was then mixed with chloroform (50 mL), filtered andconcentrated. The product was purified by column chromatography onsilica gel (eluted with hexanes-ethyl acetate 0-50%), which generated(21) (310 mg, 78% yield) as a gum. Preparation ofN-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-2-nitro-4-(pyridin-3-yl)aniline,(22). A mixture of 2-nitro-4-(pyridin-3-yl)aniline (21) (200.0 mg, 0.93mmol), (3-bromopropoxy)(tert-butyl)dimethylsilane (6) (471.0 mg, 1.86mmol), Cs₂CO₃ (303.0 mg, 0.93 mmol) in DMF (5 mL) was irradiated in amicrowave reactor for 1 h at 120° C. The mixture was concentrated underreduced pressure and the product was purified by column chromatographyon silica gel (eluted with 0-50% ethyl acetate in hexanes), whichgenerated (22) (116.3 mg, 32% yield) as a gum. Preparation ofN¹-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4-(pyridin-3-yl)benzene-1,2-diamine,(23). A solution ofN-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-2-nitro-4-(pyridin-3-yl)aniline(22) (116.3 mg, 0.30 mmol) in ethanol (25 mL) was purged with nitrogenfor 5 min and 10% Pd/C (116.0 mg) was added. The mixture was placedunder a positive hydrogen pressure from a hydrogen filled balloon. After18 h, the mixture was filtered through a pad of Celite®, the pad waswashed with ethanol (10 mL), concentrated under reduced pressure, whichprovided (23) (100.0 mg, 93%) as a gum. This material was used in thenext step without further purification.

Preparation of1-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-6-(pyridin-3-yl)-1,4-dihydroquinoxaline-2,3-dione,(25). To a solution ofN¹-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4-(pyridin-3-yl)benzene-1,2-diamine(23) (100.0 mg, 0.279 mmol) and triethylamine (141.6 mg, 1.39 mmol) inDCM (25 mL) cooled in an ice-bath was slowly added ethylchloro(oxo)acetate (24) (78.5 mg, 0.56 mmol). The ice-bath was removedand the mixture warmed to ambient temperature. After 1 h, the mixturewas concentrated under reduced pressure. The resulting residue wasdiluted with methanol (25 mL) and K₂CO₃ (193.5 mg, 1.40 mmol) was added.After 2 h, the resulting mixture was concentrated, mixed with chloroform(50 mL) filtered, concentrated under reduced pressure and the productwas purified by silica gel column chromatography (eluted with 0 to 5%methanol in chloroform), which produced (25) (39.0 mg, 34% yield) as agum.

Preparation of1-(3-hydroxypropyl)-6-(pyridin-3-yl)-1,4-dihydroquinoxaline-2,3-dione,13. To a solution of1-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-6-(pyridin-3-yl)-1,4-dihydroquinoxaline-2,3-dione(25) (39.0 mg, 0.095 mmol) in acetonitrile (5 mL) was added KF (5.8 mg,0.099 mmol) followed by TMSCl (10.8 mg, 0.099 mmol). After 1 h, themixture was concentrated under reduced pressure and the product purifiedby silica gel column chromatography (eluted with 0 to 5% methanol inchloroform), which produced 13 (8.0 mg, 28% yield) as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.13 (s, 1H), 8.88 (br s, 1H), 8.61 (br s,1H), 8.03 (d, J=8.1 Hz, 1H), 7.59-7.52 (m, 3H), 7.47 (d, J=2.1 Hz, 1H),4.69 (br s, 1H), 4.23-4.15 (m, 2H), 3.58-3.51 (m, 2H), 1.85-1.78 (m,2H). LC/MS: Eluent system C (retention time: 3.85 min); ESI-MS 298[M+H]⁺.

Compound 14 Synthesis ofN,N-dimethyl-3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propanamide,14

Compound 14 was synthesized as in Scheme 18.

Preparation of 3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propanoicacid, (27). To a solution of 3,4-dihydro-1H-quinoxalin-2-one (16) (100mg, 0.67 mmol) in THF at −5° C. was added sodium bicarbonate (100 mg,1.2 mmol) followed by methyl malonyl chloride (26) (100 mg, 0.73 mmol),after which the ice-salt bath was removed. After 1 h, the mixture wasconcentrated under reduced pressure and the resulting residue waspartitioned between ethyl acetate (3×15 mL) and 1M HCl solution (10 mL).The organic layer was dried over sodium sulfate, filtered andconcentrated under reduced pressure which generated a clear oil (250mg). The oil was dissolved in THF (15 mL) and treated with a 1M solutionof LiOH in water (8 mL). After overnight, the mixture was washed withchloroform, then the pH of aqueous layer was adjusted to 1.8 (pH meter)with a 1M solution of sodium bisulfate and the product was extractedwith ethyl acetate-tert-butanol solution (8:1). The ethylacetate-tert-butanol layer was dried over sodium sulfate, filtered andconcentrated under reduced pressure, which provided (27) (196 mg) as abrown oil. This material was used in the next step without furtherpurification.

Preparation ofN,N-dimethyl-3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propanamide14. In a sealable tube, a DMF (5 mL) solution of3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propanoic acid (27) (50mg, approximately ¼ of the obtained amount) was mixed with HATU (190 mg,0.50 mmol) and a 2M THF solution of dimethylamine (2.0 mL, 4.0 mmol) andthe tube was sealed. After overnight, the mixture was concentrated underreduced pressure and the product was purified by silica gel columnchromatography (eluent with chloroform-methanol 0%→9%), which provided14 (11.1 mg, 14% yield over 3 steps) as a pink solid.

¹H NMR (600 MHz, DMSO-d6) δ 10.70 (br s, 1H), 7.50 (br s, 1H), 7.22 (brs, 1H), 7.07-6.97 (m, 2H), 4.42-4.15 (m, 2H), 3.87-3.63 (m, 2H), 2.89(br s, 3H), 2.75 (br s, 3H). LC/MS: Eluent system C (retention time:4.55 min); ESI-MS 262 [M+H]⁺ and 260 [M−H]⁻.

Compound 15 Synthesis of4-(3-hydroxypropanoyl)-4,5-dihydro-1H-benzo[e][1,4]diazepin-2(3H)-one,15

Compound 15 was synthesized as in Scheme 19.

Preparation of 3-(benzyloxy)propanoyl chloride, (15). To a solution of3-(benzyloxy)propanoic acid (14) (430 mg, 2.39 mmol) in anhydrous DCM(12 mL) containing three drops of DMF and 1.8 g of powdered molecularsieves 4 Å, cooled in an ice bath, was added oxalyl chloride (1.2 g, 9.4mmol) in two portions over 3 min. After 30 minutes, the ice bath wasremoved. One hour after warming to room temperature, the mixture wasconcentrated under reduced pressure, which provided 2.3 g of the targetacyl chloride adsorbed on the molecular sieves surface, which wasassumed to contain 20 wt % of (15).

Preparation of4-(3-(benzyloxy)propanoyl)-4,5-dihydro-1H-benzo[e][1,4]diazepin-2(3H)-one,(29). An aliquot of 3-(benzyloxy)propanoyl chloride (15) adsorbed on themolecular sieves powder (430 mg, 0.43 mmol) was added to the solution of4,5-dihydro-1H-benzo[e][1,4]diazepin-2(3H)-one (28) (60 mg, 0.37 mmol)in DMF (5 mL) followed 30 minutes later by the addition of sodiumbicarbonate (100 mg, 1.2 mmol). After overnight, the mixture wasfiltered through a Celite® plug, the plug was washed with DCM. A fewdrops of acetic acid were added to the filtrate and the mixture wasconcentrated under reduced pressure. The residue was loaded on silicagel column as a solution in minimal amount of DCM and the product elutedwith a hexane-ethyl acetate 30%→80% gradient, which provided (29) (90mg, 75% yield) as an off-white powder.

Preparation of4-(3-hydroxypropanoyl)-4,5-dihydro-1H-benzo[e][1,4]diazepin-2(3H)-one,15. To a solution of4-(3-(benzyloxy)propanoyl)-4,5-dihydro-1H-benzo[e][1,4]diazepin-2(3H)-one(29) (90 mg, 0.27 mmol) in methanol (15 mL) was added palladium oncarbon (10% by weight, 55% wet, 60 mg, 0.025 mmol), the atmosphere inthe flask was substituted with hydrogen by connecting the flask to avacuum line for 5 min with subsequent purging with a hydrogen gas(repeated 3 times) and the suspension was stirred under positivehydrogen pressure (balloon). After overnight, the mixture was dilutedwith DCM (15 mL) and filtered through a Celite® plug. The catalyst andplug was washed with DCM (2×15 mL), and the filtrate was concentratedunder reduced pressure. The product was purified by a silica gel columnchromatography (eluted with ethyl acetate-methanol 2%→12%), which aftertrituration with ether and drying provided 15 (29 mg, 46% yield) as anoff-white powder.

¹H NMR (600 MHz, DMSO-d6) δ (two sets of signals in ratio 0.6:0.4)10.14-10.01 (two singlets, 1H), 7.36-7.23 (m, 2H), 7.13-6.98 (m, 2H),4.69 (s, 1.2H), 4.56 (s, 0.8H), 4.55-4.52 (m, 1H), 4.34 (s, 1.2H), 4.31(s, 0.8H), 3.63-3.57 (m, 2H), 2.52-2.45 (m, 2H). LC/MS: Eluent system C(retention time: 4.08 min); ESI-MS 235 [M+H]⁺ and 233 [M−H]⁻.

Compound 16 Synthesis of1-(3-hydroxypropanoyl)-3,4-dihydro-1H-benzo[e][1,4]diazepin-5(2H)-one,16

Compound 16 was synthesized as in Scheme 20.

Preparation of1-(3-(benzyloxy)propanoyl)-3,4-dihydro-1H-benzo[e][1,4]diazepin-5(2H)-one,(31). Treatment of 3,4-dihydro-1H-benzo[e][1,4]diazepin-5(2H)-one (30)(60 mg, 0.37 mmol) with the 3-(benzyloxy)propanoyl chloride (15) (1.2equivalents) as described for (29) in Scheme 19 provided after thecolumn chromatography (eluted with hexane-ethyl acetate 30%→100%)compound (31) (50 mg, 41% yield) as a white powder.

Preparation of1-(3-hydroxypropanoyl)-3,4-dihydro-1H-benzo[e][1,4]diazepin-5(2H)-one,16. Hydrogenolysis of1-(3-(benzyloxy)propanoyl)-3,4-dihydro-1H-benzo[e][1,4]diazepin-5(2H)-one(31) (50 mg, 0.15 mmol) as described for 15 in Scheme 19, afterpurification by a silica gel column chromatography (eluted with ethylacetate-methanol 2%→12%) provided compound 16 (12 mg, 34% yield) as agrey powder.

¹H NMR (600 MHz, DMSO-d6) δ 8.23 (t, J=5.3 Hz, 1H), 7.65-7.58 (m, 2H),7.53 (dd, J=7.3, 7.5 Hz, 1H), 7.41 (d, J=7.5 Hz, 1H), 4.53-4.44 (m, 2H),3.59-3.53 (m, 1H), 3.51-3.44 (m, 1H), 3.25-3.19 (m, 1H), 3.12 (dd,J=3.6, 12.7 Hz, 1H), 2.95-2.87 (m, 1H), 2.32-2.26 (m, 1H), 1.91-1.85 (m,1H). LC/MS: Eluent system C (retention time: 3.29 min); ESI-MS 235[M+H]⁺.

Compound 17 Synthesis of5-(3-hydroxypropanoyl)-4,5-dihydro-1H-benzo[b][1,4]diazepin-2(3H)-one,17

Compound 17 was synthesized as in Scheme 21.

Preparation of5-(3-(benzyloxy)propanoyl)-4,5-dihydro-1H-benzo[b][1,4]diazepin-2(3H)-one,(33). Treatment of 4,5-dihydro-1H-benzo[b][1,4]diazepin-2(3H)-one (32)(60 mg, 0.37 mmol) with the 3-(benzyloxy)propanoyl chloride (15) (1.2equivalents) as described for (29) in Scheme 19 provided after silicagel column chromatography (eluted with hexane-ethyl acetate 30%→100%)compound (33) (70 mg, 58% yield) as an off-white powder.

Preparation of5-(3-hydroxypropanoyl)-4,5-dihydro-1H-benzo[b][1,4]diazepin-2(3H)-one,17. Hydrogenolysis of5-(3-(benzyloxy)propanoyl)-4,5-dihydro-1H-benzo[b][1,4]diazepin-2(3H)-one(33) (70 mg, 0.21 mmol) as described for 15 in Scheme 19, provided afterpurification by a column chromatography (eluent ethyl acetate-methanol2%→12%) compound 17 (26 mg, 74% yield) as a grey powder.

¹H NMR (600 MHz, DMSO-d6) δ 9.77 (s, 1H), 7.45-7.35 (m, 2H), 7.27-7.11(m, 2H), 4.71 (br s, 1H), 4.47 (br s, 1H), 3.60-3.45 (m, 2H), 3.44-3.30(m, 2H), 2.33-2.23 (m, 2H), 1.90-1.83 (m, 1H). LC/MS: Eluent system C(retention time: 4.22 min); ESI-MS 235.2 [M+H]⁺ and 233.0 [M−H]⁻.

Compound 18 Synthesis of6,7-difluoro-4-(3-hydroxypropanoyl)-3,4-dihydroquinoxalin-2(1H)-one, 18

Compound 18 was synthesized as in Scheme 22.

Preparation of4-(3-(benzyloxy)propanoyl)-6,7-difluoro-3,4-dihydroquinoxalin-2(1H)-one,(34). Treatment of 6,7-difluoro-3,4-dihydroquinoxalin-2(1H)-one (33) (90mg, 0.49 mmol) with the 3-(benzyloxy)propanoyl chloride (15) (20 wt %adsorbed on molecular sieves, 430 mg, 0.45 mmol) as described for (29)in Scheme 19 provided after the silica gel column chromatography (elutedwith 20%→80% ethyl acetate in hexanes) compound (34) (120 mg, 77% yield)as an off-white powder.

Preparation of6,7-difluoro-4-(3-hydroxypropanoyl)-3,4-dihydroquinoxalin-2(1H)-one, 18.Hydrogenolysis of4-(3-(benzyloxy)propanoyl)-6,7-difluoro-3,4-dihydroquinoxalin-2(1H)-one(34) (120 mg, 0.34 mmol) as described for 15 in Scheme 19 provided afterpurification by a silica gel column chromatography (eluted with ethylacetate-methanol 2%) compound 18 (48 mg, 54% yield) as a white powder.

¹H NMR (600 MHz, DMSO-d6) δ 10.75 (s, 1H), 7.80 (dd, J=8.7, 11.2 Hz,1H), 6.99 (dd, J=8.2, 10.6 Hz, 1H), 4.69 (t, J=5.0 Hz, 1H), 4.35 (s,2H), 3.72-3.65 (m, 2H), 2.71-2.65 (m, 2H). LC/MS: Eluent system C(retention time: 5.21 min); ESI-MS 257 [M+H]⁺ and 255 [M−H]⁻.

Compound 193,4-dihydro-4-(3,3,3-trifluoro-1-oxopropyl)-2(1H)-quinoxalinone, 19

Compound3,4-dihydro-4-(3,3,3-trifluoro-1-oxopropyl)-2(1H)-quinoxalinone, 19 iscommercially available (CAS #1455652-24-0) or it can be generated by oneskilled in the art using chemistry similar to that reported herein.

Compound 20 Synthesis of4-(3-hydroxybutanoyl)-3,4-dihydroquinoxalin-2(1H)-one, 20

Compound 20 was synthesized as in Scheme 23.

Preparation of methyl 3-hydroxybutanoate, (36). An emulsion ofβ-butyrolactone (1.0 g, 11.6 mmol) in aqueous solution (15 mL) of sodiumhydroxide (40 mg, 1 mmol) was stirred for 2 h upon which it wasconcentrated under reduced pressure. Methanol (5 mL) was added to theresidue, followed by concentrated hydrochloric acid (0.2 mL, 2.3 mmol).After overnight at ambient temperature, the mixture was concentratedunder reduced pressure and the residue was partitioned between ethylacetate and 2% aqueous sodium bicarbonate solution. The separatedorganic layer was dried over sodium sulfate, filtered and concentratedunder reduced pressure, which provided (36) (550 mg, maximum 4.7 mmol).This material was used in the next step without further purification.

Preparation of methyl 3-(benzyloxy)butanoate, (37). Methyl3-hydroxybutanoate (36) (550 mg, maximum 4.7 mmol) was dissolved in DCM(4 mL) and then hexane (4 mL) was added to the solution followed by thebenzyl 2,2,2-trichloroacetamidate (2.5 g, 9.9 mmol) andtrifluoromethanesulfonic acid (0.4 g, 2.7 mmol). After overnight atambient temperature, the reaction mixture was diluted with ethyl acetate(10 mL), washed with 5% sodium bicarbonate solution, then dried overanhydrous sodium sulfate, filtered and concentrated under reducedpressure, which provided (37) (2.6 g). This material was used in thenext step without further purification.

Preparation of 3-(benzyloxy)butanoic acid, (38). Methyl3-(benzyloxy)butanoate (37) (2.6 g) dissolved in THF (10 mL) was mixedwith a 1M solution of LiOH (8 mL). After overnight, the resultingmixture was washed with chloroform (30 mL) and then a 1M solution ofsodium bisulfate was added to the separated aqueous layer to adjust pHof the solution to 2. The resulting suspension was extracted with ethylacetate (3×20 mL). The combined organic layer was dried with sodiumsulfate, filtered and concentrated under reduced pressure, whichprovided (38) (1.3 g that was approximately 60% pure based on the LC/MSanalysis @254 nm). LC/MS: Eluent system B (retention time: 5.24 min);ESI-MS 195 [M+H]⁺ and 193 [M−H]⁻. This material was used in the nextstep without further purification.

Preparation of 3-(benzyloxy)butanoyl chloride (39).3-(Benzyloxy)butanoic acid (38) (550 mg, 1.5 mmol) was converted into acorresponding acid chloride (39) following the procedure described for(15) in Scheme 19. The product (39) was used directly in the next stepwithout further purification.

Preparation of4-(3-(benzyloxy)butanoyl)-3,4-dihydroquinoxalin-2(1H)-one, (40). Asdescribed for (29) in Scheme 19, 3-(benzyloxy)butanoyl chloride (39)(1.5 mmol) was reacted with 3,4-dihydro-1H-quinoxalin-2-one (16) (150mg, 1.0 mmol) in DMF (8 mL) in the presence of sodium bicarbonate (160mg, 2 mmol). Purification of the product by silica gel columnchromatography (eluted with hexane-ethyl acetate 20%-80%) provided (40)(70 mg, 22% yield) as white powder.

Preparation of 4-(3-hydroxybutanoyl)-3,4-dihydroquinoxalin-2(1H)-one,20. Hydrogenolysis of4-(3-(benzyloxy)butanoyl)-3,4-dihydroquinoxalin-2(1H)-one (40) (70 mg,0.21 mmol) following the procedure described for 15 in Scheme 19provided after silica gel column chromatography (eluted withhexane-ethyl acetate 35%-100%) compound 20 (17 mg, 33% yield) as whitepowder.

¹H NMR (600 MHz, DMSO-d6) δ 10.67 (s, 1H), 7.54 (br s, 1H), 7.23-7.18(m, 1H), 7.05 (t, J=7.6 Hz, 1H), 7.02 (d, J=7.9 Hz, 1H), 4.71 (d, J=4.7Hz, 1H), 4.38 (d, J=16.2 Hz, 1H), 4.32 (d, J=16.2 Hz, 1H), 4.10-4.04 (m,1H), 2.71-2.66 (m, 1H), 2.51-2.46 (m, 1H), 1.05 (br s, 3H). LC/MS:Eluent system C (retention time: 5.37 min); ESI-MS 235 [M+H]⁺.

Compound 21 Synthesis of4-(3-hydroxypropanoyl)-6,7-dimethyl-3,4-dihydroquinoxalin-2(1H)-one, 21

Compound 21 was synthesized as in Scheme 24.

Preparation of 2-chloro-N-(4,5-dimethyl-2-nitrophenyl)acetamide, (42).To a stirred solution of 4,5-dimethyl-2-nitroaniline (41) (500 mg, 3.01mmol) in DMF (10 mL) was added potassium carbonate (690 mg, 5 mmol) andchloroacetyl chloride (760 mg, 6.7 mmol). After overnight, the volatilecomponents were removed under reduced pressure and the resulting residuewas partitioned between chloroform and 1M hydrochloric acid. The organiclayer was separated, dried over sodium sulfate, filtered andconcentrated under reduced pressure. Recrystallization of residue frommethanol provided (42) (340 mg, 47% yield) as a fluffy orange needles.

Preparation of N-(2-amino-4,5-dimethylphenyl)-2-chloroacetamide, (43). Asuspension of iron powder (363 mg, 6.5 mmol) in an aqueous solution (10mL) of ammonium chloride (35 mg, 0.65 mmol) and acetic acid (75 mg, 1.25mmol) was stirred in a bath at 55° C. for 15 minutes, and then asolution of 2-chloro-N-(4,5-dimethyl-2-nitrophenyl)acetamide (42) (150mg, 0.62 mmol) in DMF (3 mL) was added in one portion. After 40 minutes,to the mixture was added a saturated solution of sodium bicarbonate (5mL) and resulting suspension was filtered through a Celite® plug. Ethylacetate (30 mL) was used to wash the Celite® and the combined filtratewas stirred vigorously for 5 min and then the layers were separated. Theorganic layer was dried over sodium sulfate, filtered and concentratedunder reduced pressure which provided (43) (117 mg, 89% yield).

Preparation of3-(benzyloxy)-N-(2-(2-chloroacetamido)-4,5-dimethylphenyl)propenamide,(44). Following the procedure to generate (29) in Scheme 19N-(2-amino-4,5-dimethylphenyl)-2-chloroacetamide (43) (117 mg, 0.55mmol) was treated with the 3-(benzyloxy)propanoyl chloride (15), whichprovided after the purification by silica gel column chromatography(eluent hexane-ethyl acetate 20%-100%) compound (44) (120 mg, 58% yield)as a white solid.

Preparation of4-(3-(benzyloxy)propanoyl)-6,7-dimethyl-3,4-dihydroquinoxalin-2(1H)-one,(45). To a solution of3-(benzyloxy)-N-(2-(2-chloroacetamido)-4,5-dimethylphenyl)propenamide(44) (120 mg, 0.32 mmol) in DMF (3 mL) was added sodium hydride (120 mg,3.0 mmol, 60% dispersion in oil). After 20 minutes, the mixture wastreated with 1M hydrochloric acid (5 mL) and the resulting mixture wasextracted with chloroform (3×15 mL). The combined organic layers weredrying over sodium sulfate, filtered and concentrating under reducedpressure produced (45) which was used without further purification inthe next step.

Preparation of4-(3-hydroxypropanoyl)-6,7-dimethyl-3,4-dihydroquinoxalin-2(1H)-one, 21.Hydrogenolysis of4-(3-(benzyloxy)propanoyl)-6,7-dimethyl-3,4-dihydroquinoxalin-2(1H)-one(45) as describe in the synthesis of 15 in Scheme 19 and afterpurification by silica gel column chromatography (eluted withhexane-ethyl acetate 35%-100%) provided 21 (22 mg, 28% yield over twosteps) as an off-white powder.

¹H NMR (600 MHz, DMSO-d6) δ 10.53 (s, 1H), 7.29 (br s, 1H), 6.78 (s,1H), 4.60 (t, J=5.3 Hz, 1H), 4.29 (s, 2H), 3.74-3.62 (m, 2H), 2.65 (t,J=6.1 Hz, 2H), 2.20 (s, 3H), 2.18 (s, 3H). LC/MS: Eluent system C(retention time: 7.07 min); ESI-MS 249 [M+H]⁺ and 247 [M−H]⁻ and Eluentsystem A (retention time: 5.65 min); ESI-MS 249 [M+H]⁺ and 247 [M−H]⁻.

Compound 22 Synthesis of4-(3-hydroxycyclobutanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one, 22

Compound 22 was synthesized as in Scheme 25

Preparation of methyl 3-(benzyloxy)cyclobutanecarboxylate, (47).Treatment of methyl 3-hydroxycyclobutanecarboxylate (46) (0.50 g, 3.8mmol) with the benzyl 2,2,2-trichloroacetamidate (2.5 g, 9.9 mmol) andtrifluoromethanesulfonic acid (0.28 g, 2.7 mmol) following the procedurefor (37) in Scheme 23 provided after evaporation provided (47) (3.3 g).The material was used in the next step without further purification.

Preparation of 3-(benzyloxy)cyclobutanecarboxylic acid (48).Saponification of the methyl 3-(benzyloxy)cyclobutanecarboxylate (47)(3.3 g, maximum of 3.8 mmol) following the procedure for (38) in Scheme23 after purification by silica gel column chromatography (eluent ofhexane-ethyl acetate 0%→100%) provided (48) (750 mg). The material wasused in the next step without further purification.

Preparation of 3-(benzyloxy)cyclobutanecarbonyl chloride (49).Conversion of the 3-(benzyloxy)cyclobutanecarboxylic acid (48) (400 mg,2 mmol) into (49) was achieved by following the procedure for (15) inScheme 19. The residue obtained was used immediately in the next step.

Preparation of4-(3-(benzyloxy)cyclobutanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one,(50). Treatment of 3-(benzyloxy)cyclobutanecarbonyl chloride (49) (2mmol) with 3,4-dihydro-1H-quinoxalin-2-one (16) (150 mg, 1.0 mmol)following the procedure for (29) in Scheme 19 provided afterpurification by column chromatography (eluent of hexane-ethyl acetate20%→80%) compound (50) (143 mg, 42% yield over two steps).

Preparation of4-(3-hydroxycyclobutanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one, 22.Hydrogenolysis of4-(3-(benzyloxy)cyclobutanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one(50) (143 mg, 0.42 mmol) following the procedure for 15 in Scheme 19followed by purification with silica gel column chromatography (eluentethyl acetate-methanol 1%→5%) provided 22 (90 mg, 86% yield) as acolorless solidified foam.

¹H NMR (600 MHz, DMSO-d6) δ 10.67 (s, 1H), 7.40-7.15 (m, 2H), 7.07-7.03(m, 1H), 7.01 (d, J=7.8 Hz, 1H), 5.10 (d, J=6.7 Hz, 1H), 4.30 (br s,2H), 3.88 (br s, 1H), 2.99-2.92 (m, 1H), 2.39-2.13 (m, 2H), 1.99-1.92(m, 2H). LC/MS: Eluent system C (retention time: 5.49 min); ESI-MS 247[M+H]⁺.

Compound 23 Synthesis of2-hydroxy-N-(3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl)acetamide,23

Compound 23 was synthesized as in Scheme 26.

Preparation of 4-(3-azidopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one(51). A mixture of 4-(3-bromopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one(11) (500 mg, 1.77 mmol) and sodium azide (620 mg, 9.5 mmol) in water(10 mL) was heated at 115° C. using microwave irradiation for 45minutes. After cooling to ambient temperature, the mixture was extractedwith ethyl acetate (2×20 mL). The combined organic layer was dried oversodium sulfate, filtered and concentrated under reduced pressure, whichprovided (51) (250 mg, 57% yield) as a white powder.

Preparation of 4-(3-aminopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one,(52). Hydrogenolysis of4-(3-azidopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one (51) (250 mg, 1.02mmol) as described for 15 in Scheme 19 provided (52) in quantitativeyield (240 mg). This material was used without further purification.

Preparation of2-hydroxy-N-(3-oxo-3-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl)acetamide,23. To a solution of4-(3-aminopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one (52) (160 mg, 0.73mmol), 2-hydroxyacetic acid (250 mg, 3.3 mmol) in DMF (8 mL) was addedHATU (380 mg, 1 mmol) followed by DIPEA (500 mg, 3.9 mmol). After 1 h,the mixture was concentrated under reduced pressure and the productpurified by column chromatography (eluted with ethyl acetate-methanol1%→8%) provided 23 (186 mg, 92% yield) as a white crystalline powder.

¹H NMR (600 MHz, DMSO-d6) δ 10.69 (s, 1H), 7.77 (t, J=6.0 Hz, 1H), 7.67(br s, 1H), 7.20 (br s, 1H), 7.07-7.00 (m, 2H), 5.46 (br s, 1H), 4.34(br s, 2H), 3.75 (d, J=5.3 Hz, 2H), 2.71 (br s, 2H), 1.29-1.23 (m, 2H).LC/MS: Eluent system C (retention time: 4.03 min); ESI-MS 278 [M+H]⁺ and276 [M−H]⁻.

Compound 24 Synthesis of4-(3-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one,24

Compound 24 was synthesized as in Scheme 27.

Preparation of4-(3-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)amino)propanoyl)-3,4-dihydroquinoxalin-2(1H)-one,24. A solution of the4-(3-aminopropanoyl)-3,4-dihydroquinoxalin-2(1H)-one (52) (80 mg, 0.36mmol) in methanol (10 mL) was added into a 30 mL microwave vial equippedwith a magnetic stir bar and containing 4-chloro-7-nitrobenzofurazan(53) (80 mg, 0.4 mmol) and potassium carbonate (100 mg, 0.72 mmol). Themixture was subjected to microwave irradiation with the temperature setat 80° C. for 4 h. The resulting mixture was filtered, and the filterwashed with ethyl acetate, and the combined filtrate was concentratedunder reduced pressure. The product was purified by silica gel columnchromatography (eluted with ethyl acetate) provided 24 (21.8 mg, 16%yield) as a yellow powder.

¹H NMR (600 MHz, DMSO-d6) δ 10.65 (s, 1H), 9.38 (br s, 1H), 8.48 (br s,1H), 7.50 (br s, 1H), 7.17 (br s, 1H), 7.06-6.99 (m, 1H), 6.96 (d, J=7.8Hz, 1H), 6.38 (br s, 1H), 4.35 (s, 2H), 3.72 (br s, 2H), 3.04 (br s,2H). LC/MS: Eluent system C (retention time: 7.69 min); ESI-MS 383[M+H]⁺ and 381.0 [M−H]⁻. Eluent system B (retention time: 4.38 min);ESI-MS 383 [M+H]⁺ and 381.0 [M−H]⁻.

Compound 25 3,4-dihydro-β,3-dioxo-1(2H)-quinoxalinepropanenitrile, 25

Compound 3,4-dihydro-0,3-dioxo-1(2H)-quinoxalinepropanenitrile, 25 iscommercially available (CAS #1042790-95-3) or it can be generated by oneskilled in the art using chemistry similar to that reported herein.

Compound 26N-[3-(3,4-dihydro-3-oxo-1(2H)-quinoxalinyl)-3-oxopropyl]-acetamide, 26

CompoundN-[3-(3,4-dihydro-3-oxo-1(2H)-quinoxalinyl)-3-oxopropyl]-acetamide, 26is commercially available (CAS #1281336-03-5) or it can be generated byone skilled in the art using chemistry similar to that reported herein.

Compound 27 Synthesis of1-(3-hydroxypropyl)-6-methyl-1,4-dihydroquinoxaline-2,3-dione, 27

Compound 27 was synthesized as in Scheme 28.

Preparation of 3-(4-methyl-2-nitroanilino)propan-1-ol, (55). A mixtureof 1-fluoro-4-methyl-2-nitrobenzene (54) (0.25 g, 1.6 mmol),3-aminopropan-1-ol (242.0 mg, 3.22 mmol) and K₂CO₃ (276.0 mg, 1.99 mmol)in DMF (5 mL) was stirred at room temperature. After overnight, themixture was concentrated under reduced pressure, diluted with chloroform(5 mL), adsorbed onto silica gel (1 g) and the product purified bycolumn chromatography on silica gel (eluted with 0-5% methanol inchloroform), which generated (55) (0.32 g, 95% yield) as a white solid.

Preparation of 3-(2-amino-4-methylanilino)propan-1-ol, (56). A solutionof 3-(4-methyl-2-nitroanilino)propan-1-ol (55) (0.32 g, 1.5 mmol) inethanol (50 mL) was purged with nitrogen for 5 min and 10% Pd/C (0.32 g)was added. The mixture was stirred under positive hydrogen pressure(balloon) at room temperature for 18 h. The mixture was then filteredthrough a pad of Celite®, the pad was washed with ethanol (15 mL) andthe combined filtrate was concentrated under reduced pressure. Theproduct was purified by chromatography on silica gel (eluted with 0-10%methanol in chloroform), which provided (56) (0.16 g, 59% yield) as agum.

Preparation of1-(3-hydroxypropyl)-6-methyl-1,4-dihydroquinoxaline-2,3-dione, 27. To asolution of 3-(2-amino-4-methylanilino)propan-1-ol (56) (0.16 g, 0.89mmol) and triethylamine (0.51 g, 5.0 mmol) cooled in an ice bath, wasslowly added ethyl chloro(oxo)acetate (0.37 g, 2.7 mmol). The ice bathwas removed, and after 18 h at ambient temperature, the mixture wasconcentrated under reduced pressure. The resulting residue was dilutedwith methanol (5 mL) and K₂CO₃ (0.14 g, 1.0 mmol) was added. After 18 h,the mixture was concentrated under reduced pressure. The residue wasadsorbed onto silica gel (1 g) and the product was purified by columnchromatography on silica gel (eluted with 0-5% methanol in chloroform),which generated 27 (31.0 mg, 15% yield) as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 11.98 (br s, 1H), 7.30 (d, J=8.6 Hz, 1H),7.02 (dd, J=1.0, 8.6 Hz, 1H), 6.99 (d, J=1.0 Hz, 1H), 4.64 (t, J=5.1 Hz,1H), 4.15-4.10 (m, 2H), 3.53-3.49 (m, 2H), 2.30 (s, 3H), 1.79-1.74 (m,2H). LC/MS: Eluent system C (retention time: 6.10 min); ESI-MS 235[M+H]⁺.

Compound 28 Synthesis of1-(3-hydroxypropyl)-7-(pyridin-3-yl)-1,4-dihydroquinoxaline-2,3-dione,28

Compound 28 was synthesized as in Scheme 29.

Preparation of di-tert-butyl[2-nitro-4-(pyridin-3-yl)phenyl]-2-imidodicarbonate, (58). A mixture of2-nitro-4-(pyridin-3-yl)aniline (57) (220.0 mg, 1.02 mmol), Boc₂O (0.54g, 2.5 mmol), DMAP (24.0 mg, 0.2 mmol) and DCM (25 mL) was stirredovernight at room temperature. After which it was concentrated underreduced pressure, which generated (58) (0.42 g, 98% yield) as a gum.This material was used in the next step without further purification.

Preparation of tert-butyl [2-nitro-4-(pyridin-3-yl)phenyl]carbamate,(59). To a solution di-tert-butyl[2-nitro-4-(pyridin-3-yl)phenyl]-2-imidodicarbonate (58) (0.42 g, 1.0mmol) in DCM (25 mL) was added 1,2-diaminoethane (0.25 mL, 3.75 mmol).After overnight, it was then concentrated under reduced pressure and theproduct was purified by column chromatography on silica gel (eluted with0 to 50% ethyl acetate in hexanes), which generated (59) (0.29 g, 90%yield) as a gum. This material was used in the next step without furtherpurification.

Preparation of tert-butyl [2-amino-4-(pyridin-3-yl)phenyl]carbamate,(60). A solution of tert-butyl [2-nitro-4-(pyridin-3-yl)phenyl]carbamate(59) (0.29 g, 0.91 mmol) in ethanol (50 mL) was purged with nitrogen for5 min and 10% Pd/C (0.029 g) was added. The mixture was stirred underpositive hydrogen pressure (balloon) at room temperature for 18 h, afterwhich it was filtered through a pad of Celite®, and concentrated underreduced pressure which provided (60) (0.25 g, 96% yield) as a gum. Thismaterial was used in the next step without further purification.

Preparation of tert-butyl{2-[(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)amino]-4-(pyridin-3-yl)phenyl}carbamate,(61). To a solution of tert-butyl[2-amino-4-(pyridin-3-yl)phenyl]carbamate (60) (0.25 g, 0.88 mmol) inDMF (5 mL) was added NaH (108 mg, 2.7 mmol, 60% in oil). After 10 min,(3-bromopropoxy)(tert-butyl)dimethylsilane (6) (228.0 mg, 0.9 mmol) wasadded. After 1 h, the mixture was concentrated under reduced pressureand the product purified by column chromatography on silica gel (elutedwith 0-50% ethyl acetate in hexanes), which generated (61) (0.34 g, 85%yield) as a gum.

Preparation of tert-butyl{2-[(3-{hydroxy}propyl)amino]-4-(pyridin-3-yl)phenyl}carbamate (62). Toa solution of tert-butyl{2-[(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)amino]-4-(pyridin-3-yl)phenyl}carbamate(61) (0.34 g, 0.75 mmol) in acetonitrile (5 mL) was added KF (87.0 mg,1.5 mmol) followed by TMSCl (271.0 mg, 2.5 mmol). After 1 h, the mixturewas concentrated under reduced pressure and the product was purified bycolumn chromatography on silica gel (eluted with 0-5% methanol inchloroform), which produced (62) (178.5 mg, 70% yield) as a gum.

Preparation of3-[2-amino-5-(pyridin-3-yl)anilino]propan-1-ol-hydrochloride, (63). To asolution of tert-butyl{2-[(3-{hydroxy}propyl)amino]-4-(pyridin-3-yl)phenyl}carbamate (62)(178.5 mg, 0.52 mmol) in DCM was added HCl (1 mL, 4M in dioxane). Afterovernight, the mixture was concentrated under reduced pressure and theresidue was suspended in ether (10 mL) which after filtering and dryingproduced (63) (176 mg, 99% yield) as an off-white solid.

Preparation of1-(3-hydroxypropyl)-7-(pyridin-3-yl)-1,4-dihydroquinoxaline-2,3-dione,28. To a solution of3-[2-amino-5-(pyridin-3-yl)anilino]propan-1-ol-hydrochloride (63) (0.176g, 0.499 mmol) and triethylamine (1.0 g, 10.0 mmol) in DCM (20 mL)cooled in an ice bath was slowly added ethyl chloro(oxo)acetate (0.20 g,1.5 mmol). The ice bath was then removed and after 1 h at ambienttemperature the mixture was concentrated under reduced pressure. Theresulting residue was diluted with methanol (5 mL), and K₂CO₃ (0.21 g,1.5 mmol) was added. After 18 h, the mixture was concentrated underreduced pressure and then adsorbed onto silica gel (1 g). The productwas purified by column chromatography on silica gel (eluted with 0-5%methanol in chloroform), which provided 28 (59.0 mg, 40% yield) as awhite solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.14 (s, 1H), 8.87 (br s, 1H), 8.61 (br. s,1H), 8.07 (d, J=7.5 Hz, 1H), 7.59-7.53 (m, 3H), 7.47 (d, J=1.9 Hz, 1H),4.69 (br s, 1H), 4.23-4.10 (m, 2H), 3.58-3.50 (m, 2H), 1.84-1.79 (m,2H). LC/MS: Eluent system C (retention time: 3.85 min); ESI-MS 298[M+H]⁺.

Compound 29 Synthesis of1-(3-hydroxypropyl)-6,7-dimethyl-1,4-dihydroquinoxaline-2,3-dione, 29

Compound 29 was synthesized as in Scheme 30.

Preparation ofN-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4,5-dimethyl-2-nitroaniline,(65). To a solution of 4,5-dimethyl-2-nitroaniline (64) (200 mg, 1.21mmol) in DMF (5 mL) was added NaH (96.0 mg, 2.42 mmol, 60% in oil).After 10 min, (3-bromopropoxy)(tert-butyl)dimethylsilane (6) (455.8 mg,1.799 mmol) was added dropwise. After 1 h, the mixture was concentratedunder reduced pressure and the product was purified by columnchromatography on silica gel (eluted with 0-50% ethyl acetate inhexanes), which provided (65) (0.37 g, 90% yield) as a gum.

Preparation ofN¹-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4,5-dimethylbenzene-1,2-diamine,(66). A solution ofN-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4,5-dimethyl-2-nitroaniline(3) (0.37 g, 1.1 mmol) in ethanol (50 mL) was purged with nitrogen for 5min and 10% Pd/C (0.37 g) was added. The mixture was stirred underpositive hydrogen pressure (balloon) at room temperature. After 18 h,the mixture was filtered through a pad of Celite®, the pad was washedwith ethanol (15 mL) and concentrated under reduced pressure. Theproduct was purified by chromatography on silica gel (eluted withgradient of 0-10% MeOH-chloroform), which provided (66) (0.26 g, 77%yield) as a gum.

Preparation of1-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-6,7-dimethyl-1,4-dihydroquinoxaline-2,3-dione,(67). To a solution ofN¹-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-4,5-dimethylbenzene-1,2-diamine(66) (0.26 g, 0.85 mmol) and triethylamine (0.51 g, 5.0 mmol) in DCM (20mL) cooled in an ice bath was slowly added ethyl chloro(oxo)acetate(0.37 g, 2.7 mmol). The ice bath was removed and after 18 h at ambienttemperature the mixture was concentrated under reduced pressure. Theresidue was then diluted with methanol (5 mL) and K₂CO₃ (138.0 mg, 0.99mmol) was added. After 18 h, the mixture was concentrated under reducedpressure and the residue was mixed with chloroform (15 mL) and adsorbedonto silica gel (1 g). The product was purified by column chromatographyon silica gel (eluted with 0-5% methanol in chloroform), which provided(67) (0.18 g, 60% yield) as a white solid.

Preparation of1-(3-hydroxypropyl)-6,7-dimethyl-1,4-dihydroquinoxaline-2,3-dione, 29.To a solution of1-(3-{[tert-butyl(dimethyl)silyl]oxy}propyl)-6,7-dimethyl-1,4-dihydroquinoxaline-2,3-dione(67) (0.18 g, 0.49 mmol) in acetonitrile (5 mL) was added KF (87.0 mg,1.5 mmol) followed by TMSCl (217 mg, 2.0 mmol). After 1 h, the mixturewas concentrated under reduced pressure. The product was purified bycolumn chromatography on silica gel (eluted with 0-5% methanol inchloroform), which provided 29 (53.0 mg, 42% yield) as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 11.84 (s, 1H), 7.14 (s, 1H), 6.88 (s, 1H),4.57 (t, J=5.2 Hz, 1H), 4.09-4.02 (m, 2H), 3.48-3.42 (m, 2H), 2.19 (s,3H), 2.13 (s, 3H), 1.74-1.67 (m, 2H). LC/MS: Eluent system A (retentiontime: 5.74 min); ESI-MS 249 [M+H]⁺.

Compound 30 Synthesis of5-fluoro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 30

Compound 30 was synthesized as in Scheme 31.

Preparation of 3-(3-fluoro-2-nitroanilino)propan-1-ol, (69). To asolution of 1,3-difluoro-2-nitrobenzene (68) (300 mg, 1.9 mmol) in DMF(10 mL) was added 3-aminopropan-1-ol (140 mg, 1.9 mmol) and K₂CO₃ (788mg, 5.7 mmol). After 3 h, the mixture was concentrated under reducedpressure. The resulting residue was dissolved in DCM (75 mL) and waswashed with water (2×25 mL) and brine (25 mL). The organic phase wasdried over Na₂SO₄, filtered and concentrated under reduced pressure,which generated (69) (370 mg, 91% yield) as an orange solid. Thismaterial was used in the next step without further purification.

Preparation of 3-(2-amino-3-fluoroanilino)propan-1-ol, (70). To asolution of 3-(3-fluoro-2-nitroanilino)propan-1-ol (69) (370 mg, 1.6mmol) in EtOH (10 mL) was added 10% Pd/C (35 mg). The reaction mixturewas charged with H₂ gas before it was degassed 3 times. The reactionmixture was stirred for 4 h under a H₂ atmosphere (balloon). The mixturewas then filtered through a pad of Celite®. The filtrate wasconcentrated under reduced pressure and dried which provided (70) (300mg, 99% yield) as a white solid. This material was used in the next stepwithout further purification.

Preparation of5-fluoro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 30. To asolution of 3-(2-amino-3-fluoroanilino)propan-1-ol (70) (300 mg, 1.6mmol) in DCM (20 mL) cooled in an ice bath was added Et₃N (1.3 mL, 9.5mmol), followed by slow addition of ethyl chlorooxoacetate (0.3 mL, 2.9mmol). The ice bath was then removed. After overnight at ambienttemperature, the mixture was concentrated under reduced pressure. Theresidue was dissolved in MeOH (40 mL) and to this solution was addedK₂CO₃ (150 mg, 1.1 mmol). After 4 h at room temperature, the solid wasremoved by vacuum filtration and the filtrate was concentrated underreduced pressure. The product was purified by column chromatography onsilica gel with a gradient of MeOH/CHCl₃, 0 to 20%, which generated 30(129 mg, 33% yield) as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.05 (br s, 1H), 7.28-7.23 (m, 1H),7.22-7.16 (m, 1H), 7.15-7.07 (m, 1H), 4.66 (t, J=5.3 Hz, 1H), 4.17-4.10(m, 2H), 3.52 (q, J=6.0 Hz, 2H), 1.82-1.73 (m, 2H). ¹⁹F NMR (565 MHz,DMSO-d₆) δ −129.67-−129.69 (m, 1F). LC/MS: Eluent system C (retentiontime: 4.34 min); ESI-MS 239 [M+H]⁺.

Compound 31 Synthesis of5-chloro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 31

Compound 31 was synthesized as in Scheme 32.

Preparation of 3-(3-chloro-2-nitroanilino)propan-1-ol, (72). To asolution of 1-chloro-3-fluoro-2-nitrobenzene (71) (350 mg, 2.0 mmol) inDMF (10 mL) was added 3-aminopropan-1-ol (225 mg, 3.0 mmol) and K₂CO₃(829 mg, 6.0 mmol). After 3 h, the mixture was concentrated underreduced pressure. The resulting residue was dissolved in DCM (75 mL) andwashed with water (2×25 mL) and brine (25 mL). The separated organicphase was dried over Na₂SO₄, filtered and concentrated under reducedpressure, which provided (72) (440 mg, 95% yield) as an orange oil andwas used in next step without further purification.

Preparation of 3-(2-amino-3-chloroanilino)propan-1-ol, (73). To asolution of 3-(3-chloro-2-nitroanilino)propan-1-ol (72) (220 mg, 0.95mmol) in MeOH (10 mL) was added N₂H₄.H₂O (100 mg, 2.0 mmol), followed byRaney Ni (10 mg). The reaction mixture was warmed in a 40° C. oil bathfor 1 h. After which, the mixture was filtered through a pad of Celite®.The filtrate was concentrated under reduced pressure which provided (73)(180 mg, 94% yield) as a white solid. It was used in the next stepwithout further purification.

Preparation of5-chloro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione 31. To asolution of 3-(2-amino-3-chloroanilino)propan-1-ol (73) (180 mg, 0.89mmol) in DCM (15 mL) cooled in an ice bath was added Et₃N (0.6 mL, 1.35mmol) followed by slow addition of ethyl chlorooxoacetate (0.15 mL, 1.35mmol). After overnight, the solvent was removed under reduced pressureand the resulting residue was dissolved in MeOH (10 mL). To thissolution was added K₂CO₃ (100 mg, 0.72 mmol). After 4 h, the solid wasremoved by filtration and the filtrate was concentrated under reducedpressure. The product was purified by column chromatography on silicagel with a gradient of MeOH/CHCl₃, 0 to 20%, which generated 31 (103 mg,45% yield) as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 11.40 (br s, 1H), 7.43-7.38 (m, 1H),7.34-7.28 (m, 1H), 7.21 (t, J=8.2 Hz, 1H), 4.66 (t, J=5.1 Hz, 1H),4.20-4.12 (m, 2H), 3.53 (q, J=6.0 Hz, 2H), 1.85-1.74 (m, 2H). LC/MS:Eluent system C (retention time: 5.62 min); ESI-MS 255 [M+H]⁺.

Compound 32 Synthesis of6,7-dichloro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 32

Compound 32 was synthesized as in Scheme 33.

Preparation of 3-(4,5-dichloro-2-nitroanilino)propan-1-ol, (75). To asolution of 1,2-dichloro-4-fluoro-5-nitrobenzene (74) (300 mg, 1.4 mmol)in DMF (10 mL) was added 3-aminopropan-1-ol (161 mg, 2.1 mmol) and K₂CO₃(580 mg, 4.2 mmol). After 3 h, the solvent was removed under reducedpressure. The residue was dissolved in DCM (75 mL) and washed with water(2×25 mL) and brine (25 mL). The organic phase was dried over Na₂SO₄,filtered and concentrated under reduced pressure. This provided theproduct (75) (350 mg, 94% yield) as an orange solid and it was used inthe next step without further purification.

Preparation of 3-(2-amino-4,5-dichloroanilino)propan-1-ol (76). To asolution of 3-(4,5-dichloro-2-nitroanilino)propan-1-ol (75) (200 mg,0.75 mmol) in MeOH (10 mL) was added hydrazine.hydrate (100 mg, 2.0mmol), followed by Raney Ni (10 mg). The resulting mixture was warmed inan oil bath at 40° C. for 1 h. After which the mixture was filteredthrough a pad of Celite®. The filtrate was concentrated under reducedpressure, which provided (76) (170 mg, 96% yield) as an off-white solid.It was used in the next step without further purification.

Preparation of6,7-dichloro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 32.To a solution of 3-(2-amino-4,5-dichloroanilino)propan-1-ol (76) (170mg, 0.72 mmol) in DCM (15 mL) cooled in an ice-bath was added Et₃N (0.5mL, 3.6 mmol), followed by slow addition of ethyl chlorooxoacetate (0.12mL, 1.08 mmol). The ice bath was removed. After overnight at roomtemperature, the mixture was concentrated under reduced pressure and theresidue was dissolved in MeOH (10 mL) and K₂CO₃ (100 mg, 0.72 mmol) wasadded. After 4 h, the mixture was filtered and the filtrate wasconcentrated under reduced pressure. The product was purified by columnchromatography on silica gel with a gradient of MeOH/CHCl₃, 0 to 20%,which generated 32 (25 mg, 12% yield) as a white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.14 (br s, 1H), 7.68 (s, 1H), 7.31 (s,1H), 4.67 (t, J=5.1 Hz, 1H), 4.13-4.09 (m, 2H), 3.51 (q, J=6.0 Hz, 2H),1.80-1.72 (m, 2H). LC/MS: Eluent system C (retention time: 7.75 min);ESI-MS 289 [M+H]⁺.

Compound 33 Synthesis of6-chloro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione, 33

Compound 33 was synthesized as in Scheme 34.

Preparation of 3-(4-chloro-2-nitroanilino)propan-1-ol (78). To asolution of 4-chloro-1-fluoro-2-nitrobenzene (77) (350 mg, 2.0 mmol) inDMF (10 mL) was added 3-aminopropan-1-ol (225 mg, 3.0 mmol) and K₂CO₃(829 mg, 6.0 mmol). After 3 h, the solvent was removed under reducedpressure. The resulting residue was dissolved in DCM (75 mL) and themixture was washed with water (2×25 mL) and brine (25 mL). The separatedorganic phase was dried over Na₂SO₄, filtered and concentrated underreduced pressure, which provided (23) (310 mg, 67% yield) as an orangesolid and it was used in the next step without further purification.

Preparation of 3-(2-amino-4-chloroanilino)propan-1-ol (79). To asolution of 3-(4-chloro-2-nitroanilino)propan-1-ol (78) (200 mg, 0.87mmol) in MeOH (10 mL) was added N₂H4.H₂O (100 mg, 2.0 mmol), followed byRaney Ni (10 mg). The reaction mixture was warmed in an oil bath at 40°C. for 1 h. The mixture was then filtered through a pad of Celite®. Thefiltrate was concentrated under reduced pressure, which provided (79)(165 mg, 95% yield) as an off-white solid. It was used in next stepwithout further purification.

Preparation of6-chloro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione 33. To asolution of 3-(2-amino-4-chloroanilino)propan-1-ol (79) (165 mg, 0.82mmol) in DCM (15 mL) cooled in an ice-bath was added Et₃N (0.57 mL, 4.1mmol), followed by slow addition of ethyl chlorooxoacetate (15) (0.14mL, 1.23 mmol). The ice bath was then removed. After overnight atambient temperature, the mixture was concentrated under reduced pressureand the resulting residue was dissolved in MeOH (10 mL) and K₂CO₃ (100mg, 0.72 mmol) was added. After 4 h, the mixture was filtered and thefiltrate was concentrated under reduced pressure. The product waspurified by column chromatography on silica gel with a gradient ofMeOH/CHCl₃, 0 to 20%, which generated 33 (18 mg, 9% yield) as a whitesolid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.10 (s, 1H), 7.42 (d, J=8.7 Hz, 1H), 7.24(dd, J=2.4, 8.8 Hz, 1H), 7.20 (d, J=2.4 Hz, 1H), 4.66 (t, J=5.1 Hz, 1H),4.17-4.08 (m, 2H), 3.55-3.48 (m, 2H), 1.82-1.70 (m, 2H). LC/MS: Eluentsystem C (retention time: 6.29 min); ESI-MS 255 [M+H]⁺.

Compound 34 Synthesis of1-(3-hydroxypropyl)-6-(trifluoromethyl)-1,4-dihydroquinoxaline-2,3-dione,34

Compound 34 was synthesized as in scheme 35

Preparation of 3-[2-nitro-4-(trifluoromethyl)anilino]propan-1-ol, (81).A mixture of 1-fluoro-2-nitro-4-(trifluoromethyl)benzene (80) (500.0 mg,2.39 mmol), 3-aminopropan-1-ol (360 mg, 4.78 mmol) and K₂CO₃ (991 mg,7.17 mmol) in N,N-dimethyl formamide (10 mL) was stirred overnight atroom temperature. Then 25 mL water was added and the mixture wasextracted with CHCl₃ (3×25 mL). The combined organic layer was washedwith saturated brine solution (1×25 mL), dried over anhydrous Na₂SO₄,filtered, and concentrated under reduced pressure, which generated (81)(620 mg, 98% yield) as a yellow color liquid. This material was used inthe next step without further purification.

Preparation of 3-[2-amino-4-(trifluoromethyl)anilino]propan-1-ol, (82).To a solution of 3-[2-nitro-4-(trifluoromethyl)anilino]propan-1-ol (81)(620.0 mg, 2.35 mmol) in absolute ethanol (10 mL) was added 10% Pd/C (60mg). A balloon filled with hydrogen gas was attached onto theround-bottomed flask. After overnight, the mixture was filtered througha bed of Celite® and the bed was washed with ethanol (2×10 mL).Concentration of combined filtrates under reduced pressure produced (82)(540 mg, 98% yield) as a pale brown color solid.

Preparation of1-(3-hydroxypropyl)-6-(trifluoromethyl)-1,4-dihydroquinoxaline-2,3-dione,34. To a solution of 3-[2-amino-4-(trifluoromethyl)anilino]propan-1-ol(82) (320.0 mg, 1.37 mmol) and Et₃N (0.95 mL, 6.83 mmol) in DCM (10.0mL), was added ethyl chlorooxoacetate (280.0 mg, 2.05 mmol) dropwise.After overnight at room temperature, the mixture was concentrated underreduced pressure. To the residue was added MeOH (10 mL) and K₂CO₃ (2.0g, 14.47 mmol). After 24 h, the mixture was vacuum filtered and thecollected solid was washed CHCl₃:MeOH (9:1, 3×10 mL). The combinedfiltrates were concentrated under reduced pressure. The residue wasadsorbed on silica gel (5 g) using CHCl₃:MeOH (90:10, 10 mL) and theproduct was purified by column chromatography on silica gel (Biotage®,12 g silicycle column, eluted with gradient of 0% to 20% MeOH in CHCl₃),which afforded 34 (108.4 mg, 28% yield) as an off-white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.03 (brs, 1H), 7.60 (d, J=8.8 Hz, 1H),7.52 (dd, J=1.6, 8.8 Hz, 1H), 7.48 (s, 1H), 4.67 (t, J=5.1 Hz, 1H),4.18-4.15 (m, 2H), 3.52 (q, J=6.0 Hz, 2H), 1.82-1.75 (m, 2H). ¹⁹F NMR(565 MHz, DMSO-d₆) δ −60.45 (s, 3F). LC/MS: Eluent system A (retentiontime: 5.73 min); ESI-MS: 289 [M+H]⁺.

Compound 35 Synthesis of1-(3-hydroxypropyl)-7-(trifluoromethyl)-1,4-dihydroquinoxaline-2,3-dione,35

Compound 35 was synthesized as in scheme 36.

Preparation of 3-[2-nitro-5-(trifluoromethyl)anilino]propan-1-ol (84).Following a similar procedure as described for (81) in Scheme 35 with2-fluoro-1-nitro-4-(trifluoromethyl)benzene (83) (500 mg, 2.39 mmol),3-aminopropan-1-ol (360 mg, 4.78 mmol) and K₂CO₃ (991 mg, 7.17 mmol) inN,N-dimethylformamide (10 mL), afforded (84) (595 mg, 94% yield) as ayellow color solid. This material was used in the next step withoutfurther purification.

Preparation of 3-[2-amino-5-(trifluoromethyl)anilino]propan-1-ol (85).Following a similar procedure as described for (82) in Scheme 35 with3-[2-nitro-5-(trifluoromethyl)anilino]propan-1-ol (84) (595 mg, 2.25mmol), absolute ethanol (10 mL), and 10% Pd/C (60 mg), afforded (85)(480 mg, 91% yield) as a pale brown color solid.

Preparation of1-(3-hydroxypropyl)-6-(trifluoromethyl)-1,4-dihydroquinoxaline-2,3-dione,35. Following a similar procedure as described for 34 in Scheme 35 with3-[2-amino-5-(trifluoromethyl)anilino]propan-1-ol (85) (330 mg, 1.41mmol), Et₃N (0.98 mL, 7.05 mmol), ethyl chlorooxoacetate (290 mg, 2.11mmol) in DCM (10 mL), followed by MeOH (10.0 mL) and K₂CO₃ (2.0 g, 14.47mmol). The resulting residue was adsorbed on silica gel (5 g) usingCHCl₃:MeOH (90:10, 10 mL) and the product purified by columnchromatography on silica gel (Biotage®, 12 g silicycle column, elutedwith gradient of 0% to 20% MeOH in CHCl₃), which afforded 35 (136.1 mg,34% yield) as an off-white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.30 (brs, 1H), 7.70 (s, 1H), 7.53 (dd,J=1.0, 8.4 Hz, 1H), 7.34 (d, J=8.3 Hz, 1H), 4.71 (t, J=4.5 Hz, 1H),4.20-4.17 (m, 2H), 3.51 (q, J=5.8 Hz, 2H), 1.82-1.74 (m, 2H). ¹⁹F NMR(565 MHz, DMSO-d₆) δ −59.9 (s, 3F). LC/MS: Eluent system A retentiontime: 5.65 min); ESI-MS: 289 [M+H]⁺.

Compound 36 Synthesis of7-chloro-6-fluoro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione,36

Compound 36 was synthesized as in scheme 37.

Preparation of 3-(5-chloro-4-fluoro-2-nitroanilino)propan-1-ol (87).Following a similar procedure as described for (81) in Scheme 35 with1-chloro-2,5-difluoro-4-nitrobenzene (86) (500 mg, 2.58 mmol),3-aminopropan-1-ol (388 mg, 5.17 mmol), and K₂CO₃ (1.07 g, 7.75 mmol) inN,N-dimethylformamide (10 mL) produced (87) (620 mg, 97% yield) as ayellow solid. This material was used in the next step without furtherpurification.

Preparation of 3-(2-amino-5-chloro-4-fluoroanilino)propan-1-ol (88). Toa solution of 3-(5-chloro-4-fluoro-2-nitroanilino)propan-1-ol (87) (200mg, 0.80 mmol) in methanol (10 mL) was added Raney nickel (20 mg) andhydrazine hydrate (1.0 mL). After effervescences stopped at roomtemperature, a water condenser was attached and the mixture was heatedto reflux in an oil bath. After 1 h, the reaction mixture was cooled toroom temperature and filtered through a bed of Celite® and the Celite®bed was washed with methanol (2×10 mL). The combined filtrates wereconcentrated under reduced pressure, which generated (88) (175.9 mg,quantitative yield) as a pale brown color solid. This material was usedin the next step without further purification.

Preparation of7-chloro-6-fluoro-1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione,36. Following a similar procedure as described for 34 in Scheme 35 with3-(2-amino-5-chloro-4-fluoroanilino)propan-1-ol (88) (175.9 mg, 0.80mmol), Et₃N (0.56 mL, 4.02 mmol), and ethyl chlorooxoacetate (164.8 mg,1.21 mmol) in DCM (10 mL), followed by MeOH (10 mL) and K₂CO₃ (2.0 g,14.47 mmol). The resulting mixture was adsorbed on silica gel (5 g)using CHCl₃:MeOH (90:10, 10 mL) and the product purified by columnchromatography on silica gel (Biotage®, 12 g silicycle column, elutedwith gradient of 0 to 20% MeOH in CHCl₃), which afforded 36 (76.3 mg,35% yield) as a yellowish brown solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.10 (brs, 1H), 7.63 (d, J=6.8 Hz, 1H),7.14 (d, J=9.6 Hz, 1H), 4.68 (t, J=4.9 Hz, 1H), 4.12-4.10 (m, 2H), 3.51(q, J=5.7 Hz, 2H), 1.80-1.73 (m, 2H). ¹⁹F NMR (565 MHz, DMSO-d₆) δ−122.5 (s, 1F). LC/MS: Eluent system A (retention time: 5.15 min);ESI-MS: 273 [M+H]⁺.

Compounds 37 and 38 Synthesis of4-(2-hydroxycyclopentanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one, 37and 38

Compounds 37 and 38 were synthesized as in Scheme 38.

Preparation of methyl 2-(benzyloxy)cyclopentanecarboxylate, (90).Benzylation of methyl 2-hydroxycyclopentanecarboxylate (89) (0.5 g, 3.47mmol) in the presence of benzyl-2,2,2-trichloroacetamidate (2.3 g, 9.1mmol) and trifluoromethanesulfonic acid (0.2 g, 1.3 mmol) wasaccomplished by following a similar procedure to that for (37) in Scheme23, which provided (90) (2.81 g).

Preparation of 2-(benzyloxy)cyclopentanecarboxylic acid (91).Saponification of the crude methyl 2-(benzyloxy)cyclopentanecarboxylate(90) (2.81 g) following a similar procedure as for (38) in Scheme 23,which provided after purification by silica gel column chromatography(elute with gradient of 0%→100% ethyl acetate in hexanes) compound (91)(1.7 g) as a colorless oil. This was used in the next step withoutfurther purification.

Preparation of 2-(benzyloxy)cyclopentanecarbonyl chloride, (92).Following the procedure for (15) in Scheme 19 with2-(benzyloxy)cyclopentanecarboxylic acid (91) (0.6 g, maximum 1.2 mmol)provided (92) along with molecular sieves powder. Preparation of4-(2-(benzyloxy)cyclopentanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one,(93) and (94). Following the procedure for (29) in Scheme 19 with2-(benzyloxy)cyclopentanecarbonyl chloride (92) (0.6 g, maximum 1.2mmol) and 3,4-dihydro-1H-quinoxalin-2-one (16) (160 mg, 1.09 mmol),provided after purification by silica gel column chromatography (elutedwith gradient of 20%→80% ethyl acetate in hexanes) two fractionscontaining enriched (>80% de) diastereomers (93) (98 mg, 26% yield) and(94) (30 mg, 8% yield).

Preparation of diastereomers of4-(2-hydroxycyclopentanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one 37 and38. Two diastereomers of4-(2-(benzyloxy)cyclopentanecarbonyl)-3,4-dihydroquinoxalin-2(1H)-one(93) (98 mg, 0.28 mmol) and (94) (30 mg, 0.08 mmol) were treatedseparately as described for 15 in Scheme 19. Purification of theseparate samples by silica gel column chromatography (eluent ethylacetate-methanol 0%→4%) provided 37 (60 mg, 82% yield) and 38 (14 mg,67% yield), both as white powders.

For Compound 37:

¹H NMR (600 MHz, DMSO-d6) δ 10.68 (s, 1H), 7.47 (br s, 1H), 7.21 (s,1H), 7.09-6.99 (m, 2H), 4.97 (d, J=5.0 Hz, 1H), 4.40-4.30 (m, 2H), 4.23(quintet, J=5.6 Hz, 1H), 3.17 (d, J=5.3 Hz, 2H), 3.09-3.04 (m, 1H),1.90-1.80 (m, 2H), 1.66-1.58 (m, 2H). LC/MS: Eluent system C (retentiontime: 6.74 min); ESI-MS 261 [M+H]⁺.

For Compound 38:

¹H NMR (600 MHz, DMSO-d6) δ 10.60 (s, 1H), 7.60 (d, J=7.8 Hz, 1H), 7.17(br s, 1H), 7.05-6.98 (m, 2H), 4.72 (d, J=4.5 Hz, 1H), 4.33 (br s, 2H),3.30 (q, J=6.9 Hz, 1H), 2.10 (br s, 1H), 1.77-1.68 (m, 2H), 1.65-1.59(m, 2H), 1.58-1.52 (m, 1H), 1.43-1.35 (m, 1H). LC/MS: Eluent system C(retention time: 7.02 min); ESI-MS 261 [M+H]⁺.

Compound 39 Synthesis of4-oxo-4-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)butanenitrile, 39

Compound 39 was synthesized as in Scheme 39.

Preparation of4-oxo-4-(3-oxo-3,4-dihydroquinoxalin-1(2H)-yl)butanenitrile, 39. To asolution of 3-cyanopropanoic acid (95) (250 mg, 2.5 mmol) in DCM (6 mL)containing two drops of DMF and powdered molecular sieves (4 Å, 2 g)cooled in an ice bath was added oxalyl chloride (315 mg, 2.5 mmol).After stirring for 30 min the ice bath was removed. After 1 h at ambienttemperature, the mixture was diluted with DMF (8 mL) and3,4-dihydro-1H-quinoxalin-2-one (16) (150 mg, 1.0 mmol) was added,followed by sodium bicarbonate (500 mg, 6 mmol). After overnight at roomtemperature the mixture was filtered, and to the filtrate was added afew drops of acetic acid and then the mixture was concentrated underreduced pressure. Purification of the product was accomplished by silicagel column chromatography (eluting with a gradient of 25%→100% ethylacetate in hexanes), which provided 39 (215 mg, 94% yield) as a whitepowder.

¹H NMR (600 MHz, DMSO-d6) δ 10.71 (s, 1H), 7.54 (br s, 1H), 7.23 (br s,1H), 7.09-7.01 (m, 2H), 4.36 (br s, 2H), 2.91 (br s, 2H), 2.66 (t, J=6.8Hz, 2H). LC/MS: Eluent system C (retention time: 4.71 min); ESI-MS 230[M+H]⁺ and 228 [M−H]⁻.

Compound 40 3,4-dihydro-4-(4-hydroxy-1-oxobutyl)-2(1H)-quinoxalinone, 40

Compound 3,4-dihydro-4-(4-hydroxy-1-oxobutyl)-2(1H)-quinoxalinone, 40 iscommercially available (CAS #1512680-56-6) or it can be generated by oneskilled in the art using chemistry similar to that reported herein.

Compound 41 Synthesis of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl 2-methylpropanoate,41

Compound 41 was synthesized as in Scheme 40.

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl2-methylpropanoate, 41. To a solution of1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione 1 (75 mg, 0.34mmol) and isobutyric acid (0.046 mL, 0.51 mmol) in DCM (8 mL) was addedEDCI (104 mg, 0.54 mmol) and DMAP (125 mg, 1.02 mmol). After overnight,the mixture was diluted with DCM (15 mL) and water (5 mL). The twolayers were separated, and the organic phase was washed with brine (10mL), dried over Na₂SO₄, filtered and concentrated under reducedpressure. The product was purified by column chromatography on silicagel with a gradient of EtOAc/hexanes, 50 to 100%, which generated 41 (54mg, 55% yield) as a white solid.

¹H NMR (600 MHz, CDCl₃) δ 10.89-10.61 (br.s, 1H), 7.40-7.21 (m, 4H),4.40-4.31 (m, 2H), 4.25 (t, J=6.0 Hz, 2H), 2.62 (t, J=7.0 Hz, 1H),2.20-2.13 (m, 2H), 1.24 (d, J=7.0 Hz, 6H). LC/MS: Eluent system A(retention time: 6.78 min); ESI-MS 291 [M+H]⁺.

Compound 42 Synthesis of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl L-valinatehydrochloride salt, 42

Compound 42 was synthesized as in Scheme 41.

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propylN-(tert-butoxycarbonyl)-L-valinate (96). To a solution of1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione 1 (75 mg, 0.34mmol) and (S)-2-(Boc-amino)-3-methylbutyric acid (Boc-Val-OH) (81 mg,0.37 mmol) in DMF was added EDCI (98 mg, 0.51 mmol) and DMAP (1 mg).After 24 h, more DMAP (42 mg, 0.34 mmol) was added and after anadditional 24 h the reaction mixture was then diluted with DCM (30 mL)and the mixture was washed with water (3×5 mL) and brine (10 mL). Theorganic phase was dried over Na₂SO₄, filtered and concentrated underreduced pressure. The product was purified by column chromatography onsilica gel with a gradient of MeOH/CHCl₃, 0 to 5%, which generated (96)(80 mg, 56% yield) as a white solid.

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propylL-valinate hydrochloride salt, 42. To a solution of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propylN-(tert-butoxycarbonyl)-L-valinate (96) (80 mg, 0.19 mmol) in DCM (3 mL)was added 4N HCl solution in dioxane (2 mL, 8 mmol). After overnight,the mixture was concentrated under reduced pressure and the residue wasco-evaporated with DCM/CH₃CN (1:1, 2×10 mL). The solid was collected byfiltration and dried, which provided 42 (60 mg, 89% yield) as a whitesolid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.06 (br s, 1H), 8.32 (br s, 3H), 7.45-7.41(m, 1H), 7.25-7.14 (m, 3H), 4.37-4.31 (m, 1H), 4.29-4.18 (m, 3H), 3.93(d, J=4.5 Hz, 1H), 2.21-2.11 (m, 1H), 2.06-1.96 (m, 2H), 1.01 (d, J=7.0Hz, 3H), 0.98 (d, J=6.9 Hz, 3H). LC/MS: Eluent system A (retention time:4.09 min); ESI-MS 320 [M+H]⁺.

Compound 43 Synthesis of propan-2-yl4-(3-hydroxypropyl)-2,3-dioxo-3,4-dihydroquinoxaline-1(2H)-carboxylate,43

Compound 43 was synthesized as in Scheme 42.

Preparation of propan-2-yl4-(3-hydroxypropyl)-2,3-dioxo-3,4-dihydroquinoxaline-1(2H)-carboxylate43. To a solution of1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione 1 (75 mg, 0.34mmol) in DCM (5 mL) cooled in an ice-bath was added Et₃N (0.14 mL, 1.02mmol), followed by isopropyl chloroformate (0.37 mL, 0.37 mmol). The icebath was then removed. After 3 days at ambient temperature, the reactionmixture was diluted with DCM (15 mL) and water (10 mL) was added. Thetwo layers were separated. The aqueous layer was extracted with DCM(2×15 mL). The combined organic phase was washed with brine, dried overNa₂SO₄, filtered and concentrated under reduced pressure. The productwas purified by column chromatography on silica gel with a gradient ofMeOH/CHCl₃, 0 to 10%, which generated 43 (29 mg, 28% yield) as a whitesolid.

¹H NMR (600 MHz, CDCl₃) δ 7.86 (dd, J=1.5, 7.9 Hz, 1H), 7.64 (ddd,J=1.5, 7.2, 8.6 Hz, 1H), 7.51-7.42 (m, 2H), 5.07 (spt, J=6.3 Hz, 1H),4.52 (t, J=6.4 Hz, 2H), 3.66-3.56 (m, 2H), 2.98 (t, J=6.6 Hz, 1H),2.13-2.03 (m, 2H), 1.44 (d, J=6.3 Hz, 6H). LC/MS: Eluent system A(retention time: 7.17 min).

Compound 44 and 45 Synthesis of[4-(3-hydroxypropyl)-2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl]methylmethyl carbonate, 44 and{[4-(3-hydroxypropyl)-3-oxo-3,4-dihydroquinoxalin-2-yl]oxy}methyl methylcarbonate, 45

Compounds 44 and 45 were synthesized as in Scheme 43.

Preparation of[4-(3-hydroxypropyl)-2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl]methylmethyl carbonate, 14 and{[4-(3-hydroxypropyl)-3-oxo-3,4-dihydroquinoxalin-2-yl]oxy}methyl methylcarbonate, 15. To a solution of1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione 1 (75 mg, 0.34mmol) in DMF (5 mL) was added chloromethyl methyl carbonate (64 mg, 0.51mmol), K₂CO₃ (94 mg, 0.68 mmol) and KI (56 mg, 0.34 mmol). After 48 h,another portion of chloromethyl methyl carbonate (64 mg, 0.51 mmol),K₂CO₃ (94 mg, 0.68 mmol) and KI (56 mg, 0.34 mmol) were added. Afteranother 48 h, the mixture was diluted with EtOAc (50 mL) and washed withwater (3×10 mL) and brine (10 mL). The organic phase was dried overNa₂SO₄, filtered and concentrated under reduced pressure. The productswere purified by column chromatography on silica gel with a gradient ofMeOH/CHCl₃, 0 to 5%, which provided 44 (13 mg, 12% yield) as a whitesolid and then after continuing to eluent from 5% to 10% MeOH/CHCl₃provided 45 (18 mg, 17% yield) as a white solid.

Compound 44: ¹H NMR (600 MHz, DMSO-d₆) δ 7.65-7.57 (m, 2H), 7.54 (ddd,J=1.5, 7.1, 8.4 Hz, 1H), 7.40-7.33 (m, 1H), 6.10 (s, 2H), 4.68 (t, J=5.3Hz, 1H), 4.33-4.25 (m, 2H), 3.79 (s, 3H), 3.53 (q, J=5.8 Hz, 2H),1.84-1.78 (m, 2H). LC/MS: Eluent system A (retention time: 6.42 min);ESI-MS 309 [M+H]⁺.

Compound 45: ¹H NMR (600 MHz, DMSO-d₆) δ 7.52-7.48 (m, 2H), 7.37-7.26(m, 1H), 7.31-7.26 (m, 1H), 6.18 (s, 2H), 4.67 (s, 1H), 4.17 (dd, J=6.8,8.7 Hz, 2H), 3.76 (s, 3H), 3.54 (q, J=6.0 Hz, 2H), 1.85-1.75 (m, 2H)LC/MS: Eluent system A (retention time: 4.96 min); ESI-MS 309 [M+H]⁺.

Compound 46 Synthesis of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl 2-methylbenzoate, 46

Compound 46 was synthesized as in Scheme 44.

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl2-methylbenzoate, 46. To a stirred suspension of1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione 1 (30 mg, 0.13 mmol) inDCM (10 mL) was added 2-methylbenzoyl chloride (50 mg, 0.32 mmol, 2.5equivalents) followed by triethylamine (100 mg, 0.99 mmol). Afterovernight, the mixture was concentrated under reduced pressure, then theresidue was dissolved in a minimal amount of DCM and loaded on silicacolumn through a silica plug. Purification by column chromatography(eluted with a gradient of 0%→8% methanol in chloroform) provided 46 (14mg, 32% yield) as white powder.

¹H NMR (600 MHz, DMSO-d6) δ 12.01 (s, 1H), 7.79 (dd, J=1.3, 7.8 Hz, 1H),7.49 (ddd, J=1.5, 7.5, 7.6 Hz, 1H), 7.46-7.43 (m, 1H), 7.33 (d, J=7.7Hz, 1H), 7.31 (t, J=7.5 Hz, 1H), 7.20-7.15 (m, 3H), 4.34 (t, J=6.0 Hz,2H), 4.30 (t, J=7.0 Hz, 2H), 2.51 (s, 3H), 2.10 (quintet, J=7.1 Hz, 2H).LC/MS: Eluent system D (retention time: 4.60 min); ESI-MS 339 [M+H]⁺ and337 [M−H]⁻.

Compound 47 Synthesis of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl 4-methylbenzoate, 47

Compound 47 was synthesized as in Scheme 45.

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl4-methylbenzoate, 47. Following the procedure for 46 in Scheme 44 with1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione 1 (30 mg, 0.13 mmol),4-methylbenzoyl chloride (50 mg, 0.32 mmol) and triethylamine (100 mg,0.99 mmol) provided after product purification by column chromatography(eluted with a gradient of 0%→8% methanol in chloroform), and subsequenttrituration with ether, 47 (40 mg, 91% yield) as white powder.

¹H NMR (600 MHz, DMSO-d₆) δ 12.02 (s, 1H), 7.82 (d J=8.2 Hz, 2H),7.47-7.22 (m, 1H), 7.33 (d J=8.0 Hz, 2H), 7.21-7.15 (m, 3H), 4.34 (t,J=6.1 Hz, 2H), 4.31 (t, J=7.1 Hz, 2H), 2.39 (s, 3H), 2.11 (quintet,J=6.1 Hz, 2H). LC/MS: Eluent system D (retention time: 4.81 min); ESI-MS339 [M+H]⁺ and 337 [M−H]⁻.

Compound 48 Synthesis of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl benzoate, 48

Compound 48 was synthesized as in Scheme 46.

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propylbenzoate, 48. Following the procedure for 46 in Scheme 44 with1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione 1 (30 mg, 0.13 mmol),benzoyl chloride (50 mg, 0.36 mmol) and triethylamine (100 mg, 0.99mmol) provided after purification by column chromatography (eluted witha gradient of 0%→8% methanol in chloroform), and subsequent triturationwith ether, 48 (36 mg, 85% yield) as a white powder.

¹H NMR (600 MHz, DMSO-d₆) δ 12.03 (s, 1H), 7.92 (d J=8.3 Hz, 1H),7.69-7.65 (m, 1H), 7.54-7.51 (m, 2H), 7.47-7.44 (m, 1H), 7.21-7.15 (m,3H), 4.36 (t, J=6.0 Hz, 2H), 4.31 (t, J=7.0 Hz, 2H), 2.51 (s, 3H), 2.13(quintet, J=6.8 Hz, 2H). LC/MS: Eluent system D (retention time: 3.69min); ESI-MS 325 [M+H]⁺ and 323 [M−H]⁻.

Compound 49 Synthesis of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl 2-methoxybenzoate, 49

Compound 49 was synthesized as in Scheme 47.

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl2-methoxybenzoate, 49. Following the procedure for 46 in Scheme 44 with1-(3-hydroxypropyl)quinoxaline-2,3(1H,4H)-dione 1 (30 mg, 013 mmol),2-methoxybenzoyl chloride (50 mg, 0.29 mmol) and triethylamine (100 mg,0.99 mmol) provided after purification by column chromatography (elutedwith a gradient of 0%→8% methanol in chloroform), and subsequenttrituration with ether, 49 (32 mg, 69% yield) as a white powder.

¹H NMR (600 MHz, DMSO-d₆) δ 12.02 (s, 1H), 7.66 (dd, J=1.8, 7.6 Hz, 1H),7.56 (ddd, J=1.0, 7.5, 8.3 Hz, 1H), 7.46-7.43 (m, 1H), 7.21-7.14 (m,4H), 7.03 (ddd, J=0.8, 7.3, 7.6 Hz, 1H), 4.32 (t, J=6.0 Hz, 2H), 4.28(t, J=7.0 Hz, 2H), 3.83 (s, 3H), 2.06 (quintet, J=7.2 Hz, 2H). LC/MS:Eluent system D (retention time: 2.99 min); ESI-MS 355 [M+H]⁺ and 353[M−H]⁻.

Compound 50 Synthesis of3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propyl cyclohexylcarbamate,50

Compound 50 was synthesized as in Scheme 48

Preparation of 3-(2,3-dioxo-3,4-dihydroquinoxalin-1(2H)-yl)propylcyclohexylcarbamate, 50. A mixture of1-(3-hydroxypropyl)-1,4-dihydroquinoxaline-2,3-dione 1 (40 mg, 0.18mmol), cyclohexyl isocyanate (29.6 mg, 0.24 mmol), and Et₃N (38.0 μL,0.27 mmol) in anhydrous DCM:THF (1:1, 5.0 mL) was heated in an oil bathat 60° C. After 3 h, the reaction was cooled to room temperature andconcentrated under reduced pressure. The product was purified by columnchromatography on silica gel (Biotage®, 12 g Silicycle column, elutedwith gradient of 0 to 4% methanol in chloroform), which afforded 50(45.2 mg, 72% yield) as an off-white solid.

¹H NMR (600 MHz, DMSO-d₆) δ 12.02 (s, 1H), 7.38 (dd, J=3.4, 5.8 Hz, 1H),7.22-7.16 (m, 3H), 7.11 (d, J=7.9 Hz, 1H), 4.16 (t, J=7.2 Hz, 2H), 4.03(t, J=6.1 Hz, 2H), 3.28-3.19 (m, 1H), 1.97-1.85 (m, 2H), 1.79-1.71 (m,2H), 1.71-1.63 (m, 2H), 1.61-1.47 (m, 1H), 1.31-0.97 (m, 5H). LC/MS:Eluent system D (retention time: 3.96 min); ESI-MS: 344 [M−H]⁻.

Example 2: Testing of Compounds for Reduction of Camp in the Presence ofHuman Amylin

Compounds described herein were screened using a cAMP (cyclic adenosinemonophosphate) assay in amylin receptor subtype 3 expressing cells (SeeFIG. 1). Screening of Compound 1 and Compound 10 using cAMP assays inamylin receptor subtype 3 expressing cells indicated that they wereamylin receptor antagonists. See FIG. 2.

Cell Cultures. Amylin receptor subtypes (AMY3-HEK cells) stablyexpressed in human embryonic kidney (HEK293) cell-line were generatedand characterized as described in a previous published article (Fu etal., J. Biol. Chem. 2012). The AMY3-HEK cells were grown in a 5% CO₂humidified incubator at 37° C. with DMEM, 10% FBS, and 100 μg/mL Zeocinmedium.

ELISA (enzyme-linked immunosorbent assay). Cellular cAMP levels weremeasured using a parameter cyclic AMP assay kit (R&D Systems) accordingto the manufacturer's instructions. Briefly, AMY3-HEK cells were platedon 24-well plates overnight. These cells were then incubated with orwithout the assay compounds and hAmylin for 5 min. The cells were lysedwith lysis buffer provided in the assay kit. Standard curves wereplotted using the cAMP standards provided in the ELISA kits. All sampleswere analyzed in duplicate. The plate was measured at 450 nm. Data wasplotted, and non-linear regression was fitted with four parameters usingPrism software (GraphPad Software, La Jolla, Calif.).

Compounds were assayed based on potency of reduction at 10 μM of 0.1 μMhuman Amylin (hAmylin) induced cAMP increases.

Example 3: Compound 1 and 10 Attenuated Elevations in IntracellularCalcium Evoked by Human Amylin

Kinetic Intracellular Ca²⁺ Changes. Dynamic changes of the freecytosolic Ca2+ concentration were monitored with microplate reader. Formonitor amylin induced intracellular Ca²⁺, AMY3-HEK293 cells were platedon 96 well cell culture plates and incubated at 37° C. for 12-36 h withDMEM, 10% FBS, and Zeocin medium. For Ca²⁺ changes, the AMY3-HEK293cells were washed twice with assay solution which ion content similar toextracellular brain fluid thus containing the following: 130 mM NaCl, 4mM KCl, 1 mM MgCl₂, 2 mM CaCl₂), 10 mM HEPES, and 10 mM glucose (pH7.35). Then incubated with the membrane-permeant fluorescentCa²⁺-sensitive dye Fluo-8L-AM (AAT Bioquest, Inc., Sunnyvale, Calif.),with 5 μM of the agent for 40 min at room temperature (20-23° C.) inassay solution. The test compounds were added in various concentrationsof 0.01, 0.1, 1, and 10 μM for 10 min and kinetic measured thefluorescent intensity with Varioskan-LUX microplate reader (ThermoFisher Scientific, Waltham, Mass.). Human amylin were dissolved insterile bidistilled water at 1 mM stock solution and incubated at roomtemperature for 10 min before dilution with assay solution for use at afinal concentration of 0.5 μM. Fluorescent intensity was measured atexcitation/emission wavelength of 488/514 nm, excitation bandwidth 12nm, kinetic interval 1 min, and measure time at 100 ms.

Compounds 1 and 10 were examined for their ability to alterintracellular calcium levels in AMY3-HEK cells using this method asshown in FIG. 3.

Example 4: Compound 1 and 10 Reduced Human Amylin and Amyloid BetaInduced Cytotoxicity in Neuronal Cells

MTT Cell Death Assay. The N2a (mouse) and SK-N-SH (human) neuronal cellswere seeded to 5000 cells/well in a 96-well plate in DMEM medium, 10%FBS and incubated overnight. Cells in culture medium were incubatedeither with or without the assay compounds and Aβ₁₋₄₂, 10 μM for 24-48h. At the end of treatment, 20 μL of 5 mg/mL3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT,Sigma) was added to each well and incubated at 37° C. for 3 h. Mediumwas removed, 100 μL of MTT solvent (isopropanol with 4 mM HCl) was addedto each well, and the plates were incubated for 30 min at roomtemperature on a rotating shaker. Plates were analyzed on a microplatereader at a 562 nm wavelength.

Compounds 1 and 10 were cytoprotective against amyloid beta (Abeta)toxicity in a neurons cell line (N2a) using the MTT cell assay tomeasure cell viability. See FIG. 4.

Example 5: Compound 1 and 10 Increased Hippocampal Long TermPotentiation (LTP)

Hippocampal long term potentiation (LTP) electrophysiology experiments:an in vitro cellular surrogate for memory. Brains were quickly removedfrom mice following decapitation, placed in a cold artificial cerebralspinal fluid (aCSF) on a vibratome chamber and transverse sections cutthrough the hippocampus. The aCSF contained (in mM) 124 NaCl, 3 KCl, 2.4CaCl₂), 2 MgCl₂, 1.25 NaH₂PO₄, 26 NaHCO₃ and 10 D-glucose, and wasequilibrated with 95% 02 and 5% CO₂. Hippocampal slices (400 m thick)were maintained in aCSF-filled holding chamber at room temperature forat least 1 hour and individually transferred to the submerged glassbottom recording chamber, which was constantly perfused with aCSF (2mL/min) at 30° C. Field excitatory postsynaptic potential (fEPSP) wasrecorded with a metallic (Pt/Ir) electrode (FHC, Bowdoin, Me.) from thestratum radiatum layer of Cornu ammonis 1 region of the hippocampus(CA1) area, and the Schaffer collateral afferents were stimulated with100-μs test pulses via a bipolar cluster electrode (FHC) (Kimura et al.,2012, Kimura et al. 2016). To evaluate basal synaptic transmission, weapplied different stimulation strengths (75 μA to 300 μA in steps of 25μA) and plotted the amplitudes of presynaptic fiber volleys versus thecorresponding fEPSP slopes to compare the slope of input/output (I/O)curves of fEPSP. For long-term potentiation (LTP) experiments, thestimulus strength was set to elicit 40-50% of the maximum fEPSPamplitude and test pulses were delivered to Schaffer collaterals onceevery 30 seconds. LTP was induced by 3-theta-burst stimulation (3-TBS)protocol (each burst consisted of 4 pulses at 100 Hz with a 200-msinter-burst interval). Before 3-TBS or drug application, the responseswere monitored for at least 10 minutes to ensure a stable baseline offEPSP. To determine whether the magnitude of LTP differed significantlybetween groups, average responses during the last 20-min block ofrecordings (40-60 min after TBS) were compared. Results were fromvarious treatment groups were plotted as histograms with means±standarderror (SE). Statistical analysis was performed using one-way ANOVAfollowed by post-hoc Tukey's honestly significant difference (HSD) test(for multiple comparisons) or Student's t test (for pair-wisecomparisons). All drugs and chemicals were applied directly to the slicevia bath perfusion, which allowed for a complete exchange of theperfusate in less than a minute and a half.

LTP is a cellular surrogate of memory. In brain hippocampal slices,Compound 1 application at 1 μM blocked human amylin- and amyloid beta(Aβ)-induced depression of LTP (FIGS. 5A and 5B, FIGS. 6A and 6B). Inhippocampal brain slices from transgenic AD mice (TgCRND8), LTP waschronically depressed. Application of Compound 1 increased LTP levels(FIGS. 5C and 6C) to levels close to those observed for wild type, agematched control (WT_Cont) mice. Similarly, application of Compound 10 at1 μM blocked human amylin- and amyloid beta (Aβ)-induced depression ofLTP (FIGS. 7A and 7B, FIGS. 8A and 8B). Additionally, in AD mice(TgCRND8) Compound 10 also increased LTP levels to levels observed inWT_Cont mice (FIG. 7C and FIG. 8C).

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto.

1. A method of inhibiting activity of an amylin receptor, the methodcomprising: administering to a subject in need thereof, atherapeutically effective amount of a compound of formula (I):

wherein: R is selected from the group consisting of —H, C₁-C₆-alkyl, andsubstituted C₁-C₆-alkyl; each R¹ is independently selected from thegroup consisting of H, halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl; each R² is independently selected from the groupconsisting of —H, C₁-C₆-alkyl, and substituted C₁-C₆-alkyl; each W isselected from the group consisting of —CH₂—, —CHR³— and —CR³R⁴—; R³ andR⁴ are independently selected from the group consisting of halogen,C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl, substitutedC₃-C₆-cycloalkyl, heterocyclyl, substituted heterocyclyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, fused-heterocycle,and substituted fused-heterocycle, or together R³ and R⁴ comprise acarbocycle, substituted carbocycle or oxo; m is selected from 1, 2 or 3;Q is selected from the group consisting of —CH₂—, —C(═O)—, —CH₂C(═O)—,and —CH₂CH₂—; X is present or absent, and if present is selected fromthe group consisting of —CH₂— and —C(═O)—; Y is selected from the groupconsisting of —CH₂—, and —C(═O)—; Z is selected from the groupconsisting of —OR⁵, halogen, —CN, —NR⁵R⁵, —NHC(═O)R⁵, —NHC(═O)NR⁵R⁵,aryl, and heteroaryl; and each R⁵ is independently selected from thegroup consisting of —H, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, aryl,substituted aryl, heteroaryl, and substituted heteroaryl; or anenantiomer, a mixture of enantiomers, a mixture of two or morediastereomers, a tautomer, a mixture of two or more tautomers, or anisotopic variant thereof; or a pharmaceutically acceptable salt,solvate, or hydrate thereof.
 2. The method of claim 1, wherein theamylin receptor is an AMY3 receptor.
 3. The method of claim 1, whereinthe administering is effective for treating a disease mediated throughactivity of the amylin receptor.
 4. The method of claim 3, wherein thedisease is Alzheimer's disease.
 5. The method of claim 1, wherein thecompound is of formula (II):

wherein: R is selected from the group consisting of —H and C₁-C₆-alkyl;each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl; each R² is independently selected from the group consistingof —H, and C₁-C₆-alkyl; each W is selected from the group consisting of—CH₂—, —CHR³— and —CR³R⁴—; R³ and R⁴ are independently selected from thegroup consisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, and substitutedfused-heterocycle, or together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo; m is selected from 1, 2 or 3; Q isselected from the group consisting of —CH₂—, —C(═O)— and —CH₂C(═O)—; Yis selected from the group consisting of —CH₂—, and —C(═O)—; Z isselected from the group consisting of —OR⁵, -halogen, —NR⁵R⁵,—NHC(═O)R⁵, —NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and each R⁵ isindependently selected from the group consisting of —H, C₁-C₆-alkyl,substituted C₁-C₆-alkyl, aryl, substituted aryl, heteroaryl, andsubstituted heteroaryl; or an enantiomer, a mixture of enantiomers, amixture of two or more diastereomers, a tautomer, a mixture of two ormore tautomers, or an isotopic variant thereof; or a pharmaceuticallyacceptable salt, solvate, or hydrate thereof.
 6. The method of claim 1,wherein the compound is of formula (III):

wherein: R is selected from the group consisting of —H and C₁-C₆-alkyl;each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl; each R² is independently selected from the group consistingof —H, and C₁-C₆-alkyl; each W is selected from the group consisting of—CH₂—, —CHR³— and —CR³R⁴—; R³ and R⁴ are independently selected from thegroup consisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heterocyclyl,substituted heterocyclyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, fused-heterocycle, and substitutedfused-heterocycle, or together R³ and R⁴ comprise a carbocycle,substituted carbocycle or oxo; m is selected from 1, 2 or 3; Z isselected from the group consisting of —OR⁵, halogen, —NR⁵R⁵, —NHC(═O)R⁵,—NHC(═O)NR⁵R⁵, aryl, and heteroaryl; and each R⁵ is independentlyselected from the group consisting of —H, C₁-C₆-alkyl, substitutedC₁-C₆-alkyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl; or an enantiomer, a mixture of enantiomers, a mixture of twoor more diastereomers, a tautomer, a mixture of two or more tautomers,or an isotopic variant thereof; or a pharmaceutically acceptable salt,solvate, or hydrate thereof.
 7. The method of claim 6, wherein R is —Hor —CH₃.
 8. The method of claim 6, wherein each R¹ is selected from —H,—CH₃, —CF₃, —F, —Cl and heteroaryl.
 9. The method of claim 8, whereineach R¹ is selected from —H, —CH₃, —CF₃, —F, —Cl and pyridyl.
 10. Themethod of claim 9, wherein R¹ is —H.
 11. The method of claim 6, whereinm is
 1. 12. The method of claim 6, wherein m is
 2. 13. The method ofclaim 6, wherein R³ and R⁴ are independently selected from the groupconsisting of halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl,C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl, heteroaryl, substitutedheteroaryl, fused-heterocycle, and substituted fused-heterocycle, ortogether R³ and R⁴ comprise a C₃-C₆ carbocycle, substituted C₃-C₆carbocycle or oxo.
 14. The method of claim 13, wherein R³ and R⁴ areindependently selected from the group consisting of —F, —Cl, —CH₃, ortogether R³ and R⁴ comprise a cyclopropyl, cyclobutyl, cyclopentyl oroxo.
 15. The method of claim 6, wherein W is selected from —CH₂— and—CHR³—.
 16. The method of claim 15, wherein W is —CH₂—.
 17. The methodof claim 6, wherein Z is selected from the group consisting of —OR⁵, —F,—NHC(═O)R⁵, and heteroaryl.
 18. The method of claim 17, wherein Z isselected from the group consisting of —OR⁵, and —NHC(═O)R⁵.
 19. Themethod of claim 6, wherein each R⁵ is independently selected from —H,—CH₃, —CH₂CH₃, —CH(CH₃)₂ and phenyl.
 20. The method of claim 19, whereineach R⁵ is independently selected from —H, and —CH₃.
 21. The method ofclaim 1, wherein the compound is selected from the group consisting of:


22. A compound of Formula (IV):

wherein: R is selected from the group consisting of —H and C₁-C₆-alkyl;each R¹ is independently selected from the group consisting of —H,halogen, C₁-C₆-alkyl, substituted C₁-C₆-alkyl, C₃-C₆-cycloalkyl,substituted C₃-C₆-cycloalkyl, —OR², heterocyclyl, substitutedheterocyclyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl; each R² is independently selected from the group consistingof —H, and C₁-C₆-alkyl; each W is independently selected from the groupconsisting of —CH₂—, —CHR³— and —CR³R⁴—; R³ and R⁴ are independentlyselected from the group consisting of halogen, C₁-C₆-alkyl, substitutedC₁-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,heterocyclyl, substituted heterocyclyl, aryl, substituted aryl,heteroaryl, substituted heteroaryl, fused-heterocycle, and substitutedfused-heterocycle, or together R³ and R⁴ comprise a carbocycle, orsubstituted carbocycle; Z is selected from the group consisting of —OH,—OC(═O)CH₃, —OC(═O)Ph, and —NHC(═O)CH₃; or an enantiomer, a mixture ofenantiomers, a mixture of two or more diastereomers, a tautomer, amixture of two or more tautomers, or an isotopic variant thereof; or apharmaceutically acceptable salt, solvate, or hydrate thereof; with theproviso that the compound is not:N-[3-(3,4-dihydro-3-oxo-1(2H)-quinoxalinyl)-3-oxopropyl]acetamide.23.-33. (canceled)
 34. A compound selected from the group consisting of:


35. A compound of Formula (V), Formula (VI), or Formula (VII):

wherein: R⁷ is selected from the group consisting of —CH₂OC(═O)OR⁸,—C(CH₃)HOC(═O)OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NHR⁸, —C(═O)NR⁸R⁸, andC₁-C₆-alkyl; and each R⁸ is independently selected from C₁-C₆-alkyl,substituted C₂-C₆-alkyl, C₃-C₆-cycloalkyl, substituted C₃-C₆-cycloalkyl,heterocycyl, substituted heterocycyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl, or an enantiomer, a mixture ofenantiomers, a mixture of two or more diastereomers, a tautomer, amixture of two or more tautomers, or an isotopic variant thereof; or apharmaceutically acceptable salt, solvate, or hydrate thereof.
 36. Thecompound of claim 35, wherein the compound is selected from the groupconsisting of:


37. A method of inhibiting activity of an amylin receptor, the methodcomprising: administering to a subject in need thereof, atherapeutically effective amount of a compound of claim
 22. 38. Themethod of claim 37, wherein the amylin receptor is an AMY3 receptor. 39.The method of claim 37, wherein the administering is effective fortreating a disease mediated through activity of the amylin receptor. 40.The method of claim 37, wherein the disease is Alzheimer's disease.